Method for testing sample, kit for testing gynecological cancer and precancerous lesion thereof, and pharmaceutical product

Abstract

Provided are: a simple and highly accurate test method for a sample that can be applied to the evaluation of the presence or absence of one or more species selected from the group consisting of gynecological cancer and precancerous lesions thereof, the evaluation of the prognosis of a gynecological cancer patient, or the evaluation of the degree of malignancy of gynecological cancer; and a test kit that can be used in the test method. And a pharmaceutical agent for treating or preventing gynecological cancer, which is administered to a subject detected by the examination method. A method for inspecting a sample, which comprises: a step (A) wherein the concentration of a free fatty acid in a sample derived from a subject is measured; and a step (B) in which the likelihood that the subject has a gynecological cancer is evaluated on the basis of the concentration of the free fatty acid obtained in step (A).

Description

Technical Field

[0001] The present invention relates to a method for testing a sample, a test kit for gynecological cancer and its precancerous lesions, and a pharmaceutical product. This application claims priority based on Japanese Patent Application No. 2022-128403 filed in Japan on August 10, 2022, and Japanese Patent Application No. 2023-025389 filed in Japan on February 21, 2023, and the contents thereof are incorporated herein by reference. Background Art

[0002] Ovarian cancer is a tumor that develops in the abdominal cavity, so it is difficult to show symptoms. In addition, it cannot be diagnosed pathologically through biological examination, so it is extremely difficult to detect early. In addition, there are no established serum tumor markers for early diagnosis of other gynecological cancers such as uterine cancer and cervical cancer. At present, there are many cases where they are accidentally discovered due to irregular bleeding, etc.

[0003] As an ovarian cancer tumor marker, for example, CA125 is known (for example, refer to Patent Document 1). However, the blood concentration of CA125 is not necessarily high in the case of early ovarian cancer, mucinous ovarian cancer, and ovarian clear cell carcinoma, so these cancers may not be judged as positive. In addition, the blood concentration of CA125 sometimes increases in benign diseases and inflammatory diseases, so the false positive rate of judgment based on the blood concentration of CA125 is high. Therefore, CA125 is insufficient as a tumor marker for ovarian cancer.

[0004] In recent years, as a diagnostic method for cancer, a method for detecting specific exosomes or microRNAs in peripheral blood has been proposed (for example, see Non-Patent Document 1). However, when analyzing exosomes and microRNAs from biological samples, operations such as ultracentrifugation and affinity separation are required. Therefore, the examination of them as tumor markers is complicated and time-consuming. In addition, there is a certain number of false positives, and it has not yet reached practical use. Prior art literature Patent Literature

[0005] Patent Document 1: Japanese Patent No. 6538887 Non-patent literature

[0006] Non-patent literature 1: Yu Zhang et al., Emerging Functions and Clinical Applications of Exosomal ncRNAs in Ovarian Cancer. Front. Oncol., 05 November 2021, https: / / doi.org / 10.3389 / fonc.2021.765458 Summary of the invention Problems to be solved by the invention

[0007] As described above, conventional cancer screening methods have problems in terms of simplicity and accuracy. Therefore, a method is required that can detect gynecological cancer at an early stage with high accuracy using a simple method.

[0008] Therefore, the object of the present invention is to provide a simple and highly accurate sample inspection method that can be applied to the evaluation of the presence of one or more selected from the group consisting of gynecological cancer and its precancerous lesions, the evaluation of the prognosis of gynecological cancer patients, or the evaluation of the malignancy of gynecological cancer, an inspection kit that can be used for the inspection method, and a pharmaceutical for the treatment or prevention of gynecological cancer to be administered to a subject detected by the inspection method. Means for solving problems

[0009] The present invention includes the following aspects. [1] A method for testing a sample, comprising: a step (A1) of measuring the concentration of one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, docosapentaenoic acid and arachidonic acid in a sample derived from a subject; and a step (B1) of evaluating the possibility that the subject has one or more free fatty acids selected from the group consisting of gynecological cancer and precancerous lesions thereof based on the concentration of the free fatty acids in the sample obtained in the step (A1). In the step (B1), when at least one selected from the group consisting of the following (i1), (ii1) and (iii1) is satisfied, it is evaluated that the subject has a high possibility of having one or more selected from the group consisting of gynecological cancer and precancerous lesions thereof: (i1) the concentration of at least one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid and arachidonic acid in the sample is higher than a predetermined threshold value; (ii1) the concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid in the sample is below a predetermined threshold; (iii1) The concentration ratio of docosapentaenoic acid to oleic acid is lower than a predetermined threshold value.

[0010] [2] A method for testing a sample, comprising: a step (A2) of measuring the concentration of one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid in a sample derived from a subject having gynecological cancer; and a step (B2) of evaluating the possibility of the subject having a poor prognosis based on the concentration of the free fatty acids in the sample obtained in the step (A2). In the step (B2), when at least one selected from the group consisting of the following (i2) and (ii2) is satisfied, it is evaluated that the subject has a high possibility of having a poor prognosis: (i2) the concentration of at least one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid and palmitic acid in the sample is higher than a predetermined threshold; (ii2) the concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid in the sample is lower than a predetermined threshold value.

[0011] [3] A method for testing a sample, comprising: a step (A3) of measuring the concentration of one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid in a sample derived from a subject having gynecological cancer; and a step (B3) of evaluating whether the gynecological cancer of the subject is a malignant tumor or a borderline malignant tumor based on the concentration of the free fatty acids in the sample obtained in the step (A3). In the step (B3), when at least one selected from the group consisting of the following (i3) and (ii3) is satisfied, it is evaluated that the possibility that the gynecological cancer is a malignant tumor is high: (i3) the concentration of at least one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid and palmitic acid in the sample is higher than a predetermined threshold; (ii3) the concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid in the sample is lower than a predetermined threshold value.

[0012] [4] The method for testing a sample according to [1], further comprising a step (C1), in which, based on the concentration of the free fatty acid in the sample obtained in the step (A1), the subject determined by the step (B1) to have a high probability of having one or more selected from the group consisting of gynecological cancer and its precancerous lesions is evaluated as to whether it has gynecological cancer or gynecological cancer precancerous lesions. In the step (C1), when the following (Ci) is satisfied, it is evaluated that the subject is highly likely to have gynecological cancer: (Ci) the concentration of at least one free fatty acid selected from the group consisting of dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid in the sample is lower than a predetermined threshold value.

[0013] [5] The sample inspection method according to any one of [1] to [4], wherein the gynecological cancer includes at least one selected from the group consisting of ovarian cancer, uterine cancer, and cervical cancer. [6] The sample inspection method according to [1], wherein the gynecological cancer is a stage I or stage II gynecological malignant tumor. [7] The method for inspecting a sample according to [6], wherein the step (A1) is a step of measuring the concentration of one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid and stearic acid, The free fatty acid in (i1) of the step (B1) is at least one selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid and palmitic acid, and the free fatty acid in (ii1) is at least one selected from the group consisting of vaccenic acid, arachidic acid and stearic acid. [8] The sample testing method according to any one of [1] to [4], wherein the sample is a blood sample.

[0014] [9] A test kit for gynecological cancer and its precancerous lesions, wherein the test kit comprises a reagent for measuring one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, docosapentaenoic acid and arachidonic acid, The test kit is used for at least one purpose selected from the group consisting of the following (a) to (d): (a) evaluating the possibility that the subject has one or more types of cancer selected from the group consisting of gynecological cancer and precancerous lesions thereof; (b) assessing the likelihood that a subject with gynecological cancer will have a poor prognosis; (c) whether the gynecological cancer the subject has is a malignant tumor or a borderline malignant tumor; (d) evaluating whether a subject having a high probability of having one or more selected from the group consisting of gynecological cancer and gynecological cancer precancerous lesions has gynecological cancer or gynecological cancer precancerous lesions.

[10] The gynecological cancer testing kit according to [9], wherein the testing kit is applied to the sample testing method according to any one of [1] to [4].

[0015]

[11] A pharmaceutical for treating or preventing gynecological cancer, wherein the pharmaceutical is administered to a subject assessed as having gynecological cancer by the examination method described in [1]. Effects of the Invention

[0016] According to the present invention, there can be provided a simple and highly accurate sample inspection method applicable to the evaluation of the presence or absence of gynecological cancer, the evaluation of the prognosis of gynecological cancer patients, or the evaluation of the malignancy of gynecological cancer, an inspection kit applicable to the inspection method, and a pharmaceutical product for the treatment or prevention of gynecological cancer administered to a subject determined by the inspection method. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1A This is a diagram showing the analysis results of the expression levels of fatty acid metabolic enzymes in normal ovarian tissue (Normal tissue) and ovarian cancer tissue (OV tumor tissue) in Experimental Example 1 using a heat map. Figure 1B This is a graph showing the analysis results of the expression levels of FASN and SCD1 in normal ovarian tissue (NT) and ovarian cancer tissue (Tu) in Experimental Example 1. Figure 1C This is a graph showing the analysis results of the expression levels of ELOVL1, ELOVL2, ELOVL3, ELOVL4, ELOVL5, ELOVL6, and FADS1 in normal ovarian tissue (NT) and ovarian cancer tissue (Tu) in Experimental Example 1. Figure 1D This is a graph showing the analysis results of the expression levels of ACACA and FADS2 in normal ovarian tissue (NT) and ovarian cancer tissue (Tu) in Experimental Example 1.

[0018] Figure 2A This is a graph showing the analysis results of free fatty acid concentrations in the serum of female healthy donors (Healthy donor) and ovarian cancer patients (OV ca patients) in Experimental Example 2 in the form of a heat map. Figure 2B This is a graph showing the analysis results of the concentrations of palmitoleic acid, palmitic acid, γ-linolenic acid, stearidonic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid and stearic acid in the serum of female healthy donors (HD) and ovarian cancer patients (OV) in Experimental Example 2. Figure 2C This is a graph showing the analysis results of the concentrations of arachidic acid, arachidonic acid, eicosapentaenoic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, docosapentaenoic acid, neuraminic acid and lignoceric acid in the serum of female healthy donors (HD) and ovarian cancer patients (OV) in Experimental Example 2. Figure 2D This is a diagram showing the analysis results of the free fatty acid concentration in the serum of ovarian cancer patients in Experimental Example 2 using a volcano plot. In the figure, "OV ca" and "healthy" represent ovarian cancer patients and female healthy donors, respectively.

[0019] Figure 3A This is a graph showing the correlation between the change in the expression of the fatty acid metabolic enzyme in Experimental Example 1 and the change in the fatty acid composition in Experimental Example 2. Figure 3B This is a graph showing the correlation between the change in the expression of the fatty acid metabolic enzyme in Experimental Example 1 and the change in the fatty acid composition in Experimental Example 2. Figure 3C This is a graph showing the correlation between the SCD1 gene expression level in ovarian tumor tissue in Experimental Example 3 and the fatty acid ratio (product fatty acid / substrate fatty acid ratio (palmitoleic acid / palmitic acid ratio)) corresponding to SCD1 activity in the serum of ovarian cancer patients. Figure 3D This is a graph showing the correlation between the SCD1 gene expression level in ovarian tumor tissue in Experimental Example 3 and the fatty acid ratio (product fatty acid / matrix fatty acid ratio (oleic acid / stearic acid ratio)) corresponding to SCD1 activity in the serum of ovarian cancer patients.

[0020] Figure 4A This is a graph showing the results of ROC analysis based on the serum concentrations of palmitoleic acid, linoleic acid, α-linolenic acid, palmitic acid, and oleic acid for female healthy donors and ovarian cancer patients in Experimental Example 4. Figure 4B This is a graph showing the results of ROC analysis based on the serum concentrations of vaccenic acid, eicosanoic acid, docosahexaenoic acid, adrenic acid, nervonic acid, and lignoceric acid for female healthy donors and ovarian cancer patients in Experimental Example 4.

[0021] Figure 5A This is a graph showing the results of determining whether or not the serum concentrations of 10 types of free fatty acids for healthy female donors in Experimental Example 5 are abnormal values ​​based on threshold values. Figure 5B This is a graph showing the results of determining whether or not the serum concentrations of 10 types of free fatty acids are abnormal values ​​based on threshold values ​​for patients with ovarian malignant tumors in Experimental Example 5. Figure 5C This is a graph showing the results of ROC analysis based on FFA scores for female healthy donors and ovarian malignant tumor patients in Experimental Example 5.

[0022] Fig. 6A This is a graph showing the analysis results of the serum concentrations of palmitoleic acid, linoleic acid, α-linolenic acid, and oleic acid in female healthy donors (HD), stage I and II ovarian cancer patients, and stage III and IV ovarian cancer patients in Experimental Example 6. Figure 6B The graph shows the analysis results of serum concentrations of vaccenic acid, arachidic acid, docosahexaenoic acid, and lignoceric acid in female healthy donors (HD), stage I and II ovarian cancer patients, and stage III and IV ovarian cancer patients in Experimental Example 6.

[0023] Fig. 7A This is a graph showing the results of ROC analysis based on the serum concentrations of palmitoleic acid, linoleic acid, α-linolenic acid, and oleic acid for female healthy donors and stage I or stage II ovarian cancer patients in Experimental Example 7. Figure 7B This is a graph showing the results of ROC analysis of serum concentrations of vaccenic acid, arachidic acid, docosahexaenoic acid, and lignoceric acid in female healthy donors and stage I or stage II ovarian cancer patients in Experimental Example 7.

[0024] Fig. 8A This is a graph showing the results of determining whether or not the serum concentrations of eight free fatty acids for healthy female donors in Experimental Example 8 are abnormal values ​​based on threshold values. Figure 8B This is a graph showing the results of determining whether or not the serum concentrations of eight free fatty acids for ovarian cancer patients in Experimental Example 8 are abnormal values ​​based on threshold values. Figure 8C This is a graph showing the results of ROC analysis based on FFA scores for female healthy donors (HD) and early ovarian cancer patients (stages I and II) in Experimental Example 8. Fig.8D This is a graph showing the distribution of FFA scores and CA125 blood concentrations in early ovarian cancer patients (stages I and II) in Experimental Example 8. In the graph, "Clear cell" and "Others" respectively represent ovarian clear cell cancer patients and ovarian cancer patients of tissue types other than ovarian clear cell cancer.

[0025] Fig. 9This is a graph showing the transition of the progression-free survival rate (PFS) in the FFA score high group and the FFA score low group in Experimental Example 9.

[0026] Fig. 10A This is a graph showing the results of determining whether or not the serum concentrations of 10 types of free fatty acids are abnormal values ​​based on threshold values ​​for patients with borderline ovarian malignant tumors in Experimental Example 10. Fig. 10B This is a graph showing the results of ROC analysis based on FFA scores for patients with borderline ovarian malignant tumors and patients with ovarian cancer (patients with ovarian malignant tumors) in Experimental Example 10.

[0027] Fig.11A This is a graph showing the analysis results of the concentrations of palmitoleic acid, palmitic acid, linoleic acid, α-linolenic acid, and oleic acid in the serum of female healthy donors, uterine cancer patients, and cervical cancer patients in Experimental Example 11. Fig. 11B This is a graph showing the analysis results of the concentrations of vaccenic acid, arachidic acid, and stearic acid in the serum of female healthy donors, uterine cancer patients, and cervical cancer patients in Experimental Example 11.

[0028] Fig. 12A The graph shows the analysis results of the free fatty acid concentrations of palmitoleic acid, linoleic acid, α-linolenic acid and oleic acid in the serum of female healthy donors and stage I, stage II or stage III cervical cancer patients in Experimental Example 12, as well as the ROC analysis results based on the serum concentrations. Fig. 12B The present invention is a graph showing the analysis results of the free fatty acid concentrations of vaccenic acid, arachidic acid, dihomo-γ-linolenic acid and docosahexaenoic acid in the serum of healthy female donors and stage I, II or III cervical cancer patients in Experimental Example 12, as well as the ROC analysis results based on the serum concentrations. Fig. 12C The graph shows the analysis results of serum free fatty acid concentrations of adrenic acid and docosapentaenoic acid for female healthy donors and stage I, II, or III cervical cancer patients in Experimental Example 12, and the ROC analysis results based on the serum concentrations.

[0029] Fig.13A It is a graph showing the analysis results of the concentration ratio (docosapentaenoic acid / oleic acid) in the serum of female healthy donors and stage I, stage II, or stage III cervical cancer patients in Experimental Example 13 and the ROC analysis results.

[0030] Fig. 13BThis is a graph showing the distribution of the blood concentrations of SCC markers and CEA in female healthy donors and stage I, stage II, or stage III cervical cancer patients in Experimental Example 13, and the concentration ratio (docosapentaenoic acid / oleic acid).

[0031] Fig.14A The graph shows the analysis results of serum free fatty acid concentrations of palmitoleic acid, linoleic acid, α-linolenic acid, and oleic acid in female healthy donors and patients with cervical cancer precancerous lesions in Experimental Example 14, and the ROC analysis results based on the serum concentrations. Fig. 14B The graphs show the analysis results of serum free fatty acid concentrations of arachidonic acid and arachidic acid for female healthy donors and patients with cervical cancer precancerous lesions in Experimental Example 14, and the ROC analysis results based on the serum concentrations.

[0032] Fig.15 It is a graph showing the analysis results of the free fatty acid concentrations of dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid in the serum of patients with cervical precancerous lesions (CIN) and cervical cancer patients in Experimental Example 15, and the ROC analysis results based on the serum concentrations. DETAILED DESCRIPTION

[0033] [Test sample inspection method (1)] The sample testing method of the first embodiment can be used to evaluate the possibility that a subject has gynecological cancer. The testing method includes the following steps (A1) and (B1). Step (A1): a step of measuring the concentration of one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, docosapentaenoic acid and arachidonic acid in a sample from a subject. Step (B1): a step of evaluating the possibility that the subject has one or more selected from the group consisting of gynecological cancer and precancerous lesions thereof based on the concentration of the free fatty acid in the sample obtained in step (A1).

[0034] The "subject" from which the sample for examination is derived is a human female. In this specification, gynecological cancer refers to a cancer that human males do not suffer from but human females may suffer from. In other words, gynecological cancer refers to a cancer that may occur in female-specific organs (organs that human males do not have but human females have). Examples of female-specific organs include the ovary and the uterus. As gynecological cancers, for example, ovarian cancer, uterine body cancer, and cervical cancer can be cited. In the present specification, the term "cancer" includes malignant tumors and borderline malignant tumors. Ovarian cancer can be an ovarian malignant tumor or an ovarian borderline malignant tumor. Uterine body cancer can be a uterine body malignant tumor or a borderline malignant tumor of the uterine body. Cervical cancer can be a cervical malignant tumor or a borderline malignant tumor of the cervix. The gynecological cancer detected by the inspection method of this embodiment is preferably a malignant tumor, more preferably an ovarian malignant tumor, a uterine body malignant tumor, or a cervical malignant tumor. Hereinafter, malignant tumors in gynecological cancers are also referred to as "gynecological malignant tumors". "Borderline malignant tumors" in gynecological cancers are also referred to as "gynecological borderline malignant tumors".

[0035] The stage of the gynecological cancer to be detected in the inspection method of the present embodiment is not particularly limited. In addition, the stage of cancer described in this specification is based on the TNM classification. In the inspection method of the present embodiment, it is possible to widely detect cancers ranging from early gynecological cancer (e.g., stage I, II) to stage progression (e.g., stage III, IV). The gynecological cancer to be detected may also be an early gynecological malignancy. Early gynecological malignancy refers to a gynecological malignancy of stage 0, stage I or stage II. Early gynecological malignancy is preferably a malignant tumor of stage I or stage II. As an early gynecological malignancy to be detected, an early ovarian malignancy is preferably used. The gynecological cancer to be detected may be a malignant tumor of stage III or stage IV. The gynecological cancer to be detected in the inspection method of the present embodiment may also be a gynecological malignancy with a degree of progression above stage I. Gynecological malignancies with a degree of progression above stage I include gynecological malignancies of stage I, stage II, stage III and stage IV.

[0036] In this specification, the term "precancerous lesion" refers to a tissue that is more likely to develop cancer than normal tissue. That is, a "precancerous lesion" is a tissue that is highly likely to develop into cancer. "Precancerous lesions of gynecological cancer" refers to a tissue that is highly likely to develop into "gynecological cancer". Precancerous lesions are also called intraepithelial neoplasia.

[0037] (Process (A1)) In step (A1), the concentration of one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, docosapentaenoic acid, and arachidonic acid in a sample from a subject is measured. The measurement may be in vitro. Hereinafter, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, docosapentaenoic acid and arachidonic acid are also collectively referred to as "free fatty acids of group A1".

[0038] The sample derived from the object is not particularly limited as long as the effect of the present invention is exerted, and examples thereof include biological samples such as urine, blood, saliva, sweat, and tissue pieces, and blood is preferred. Examples of blood samples include whole blood, plasma, and serum.

[0039] The method for measuring the concentration of the free fatty acids in the sample is not particularly limited. For example, the concentration of the free fatty acids in the sample can be measured using methods well known to those skilled in the art. As such a method, for example, gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, gas chromatography, etc. can be listed.

[0040] In step (A1), the number of types of free fatty acids to be measured may be one or more of the free fatty acids selected from group A1, may be two or more, may be three or more, may be four or more, may be five or more, may be six or more, may be seven or more, may be eight or more, may be nine or more, may be ten or more, may be eleven or more, may be twelve or more, may be thirteen or more, may be fourteen or more, may be fifteen. The free fatty acids to be measured may be arbitrarily selected from the free fatty acids in group A1.

[0041] The free fatty acid to be measured can be one or more (for example, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen) selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, and docosapentaenoic acid. Alternatively, the free fatty acid to be measured can be one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten) selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccinium acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, and lignoceric acid. Alternatively, the free fatty acid to be measured can be one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, or eight) selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, and lignoceric acid. Alternatively, the free fatty acid to be measured may be one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, or twelve) selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid. Alternatively, the free fatty acid to be measured can be one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, or eight) selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, and arachidic acid. Alternatively, the free fatty acid to be measured may be one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten) selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid. Alternatively, the free fatty acid to be measured may be one or more (e.g., two or more, three or more, four or more, five or more, or six) selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, arachidonic acid, and eicosanoic acid.

[0042] When the detection object is gynecological cancer, in step (A1), more than one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid can be used as the measurement object. More than 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these 14 fatty acids. When the detection object is gynecological cancer, in step (A1), the concentrations of the 14 free fatty acids are preferably measured.

[0043] When the gynecological cancer that can be used as the detection object is ovarian cancer, in step (A1), one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid and lignoceric acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these ten fatty acids. When the gynecological cancer is ovarian cancer, in step (A1), the concentrations of the ten free fatty acids are preferably measured.

[0044] In order to improve the detection sensitivity of early gynecological malignancies (e.g., stages I and II), in step (A1), one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid and lignoceric acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, or eight free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these eight fatty acids. When it is desired to detect early gynecological malignancies, it is preferred to measure the concentrations of the eight free fatty acids in step (A1). As early gynecological malignancies, for example, early ovarian malignancies can be cited.

[0045] When the gynecological cancer that can be used as the detection object is uterine body cancer or cervical cancer, in step (A1), one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, or twelve free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these twelve fatty acids. When the gynecological cancer is uterine body cancer or cervical cancer, in step (A1), the concentrations of the twelve free fatty acids are preferably measured.

[0046] When the gynecological cancer that can be used as the detection object is uterine body cancer, in step (A1), one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, or eight free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these eight fatty acids. When the gynecological cancer is uterine body cancer, in step (A1), the concentrations of the eight free fatty acids are preferably measured.

[0047] When the gynecological cancer that can be used as the detection object is cervical cancer, in step (A1), one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, or twelve free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these twelve fatty acids. When the gynecological cancer is cervical cancer, in step (A1), the concentrations of the twelve free fatty acids are preferably measured.

[0048] When the detection includes gynecological cancer precancerous lesions (e.g., cervical cancer precancerous lesions), in step (A1), the free fatty acids used for evaluation may be one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, arachidonic acid, and arachidic acid. Two or more, three or more, four or more, five or more, or six free fatty acids among the free fatty acids may be measured.

[0049] (Process (B1)) In step (B1), the possibility that the subject has gynecological cancer is evaluated based on the concentration of the free fatty acid in the sample obtained in step (A1).

[0050] In step (B1), when at least one selected from the group consisting of the following (i1) and (ii1) is satisfied, it is evaluated that the subject is highly likely to have one or more selected from the group consisting of gynecological cancer and precancerous lesions thereof.

[0051] (i1) The concentration of at least one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid and arachidonic acid in the sample is higher than a predetermined threshold value. (ii1) the concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid in the sample is lower than a predetermined threshold value. (iii1) The concentration ratio of docosapentaenoic acid to oleic acid is lower than a predetermined threshold value.

[0052] The free fatty acids described in (i1) above (palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid and arachidonic acid) are collectively referred to as "free fatty acids of group B (i1)". The free fatty acids described in (ii1) above (vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid) are collectively referred to as "free fatty acids of group B (ii1)". The "predetermined threshold value" in (i1) and (ii1) above is a value set for each free fatty acid. In the case where the free fatty acid measured in step (A1) is a free fatty acid of group B(i1), when the concentration of the free fatty acid measured in step (A1) is higher than a predetermined threshold value set for the free fatty acid, it is evaluated that the subject is likely to have one or more selected from the group consisting of gynecological cancer and its precancerous lesions. In the case where the free fatty acid measured in step (A1) is a free fatty acid of group B(ii1), when the concentration of the free fatty acid measured in step (A1) is lower than a predetermined threshold value set for the free fatty acid, it is evaluated that the subject is likely to have one or more selected from the group consisting of gynecological cancer and its precancerous lesions. When there are two or more free fatty acids measured in step (A1), these free fatty acids are classified into free fatty acids of group B (i1) and free fatty acids of group B (ii1), and whether each free fatty acid belongs to (i1) or (ii1) is confirmed. The higher the ratio of the number of free fatty acids belonging to (i1) or (ii1) relative to the number of free fatty acids measured, the higher the possibility that the subject has one or more selected from the group consisting of gynecological cancer and precancerous lesions thereof.

[0053] The predetermined threshold value of the concentration of each free fatty acid in the above (i1) and (ii1) can be a critical value obtained by obtaining the concentration of each free fatty acid from multiple samples from healthy female donors and multiple samples from patients with one or more diseases selected from a group consisting of gynecological cancer and its precancerous lesions, and performing statistical processing on the concentrations of each free fatty acid.

[0054] The predetermined threshold value of the concentration of a specific free fatty acid can be set, for example, so that the positive likelihood ratio for the specific free fatty acid becomes the maximum. The threshold value at which the positive likelihood ratio becomes the maximum can be set, for example, by performing ROC analysis. For example, for an arbitrary number of patients having one or more selected from the group consisting of gynecological cancer and precancerous lesions thereof, and a female healthy donor group, the serum concentration of the specific free fatty acid can be measured, and these serum concentrations are used to perform ROC analysis.

[0055] When the sample derived from the subject used in the step (A1) is serum, examples of the threshold values ​​of the concentrations of the free fatty acids in (i1) and (ii1) include the following threshold values, but are not limited thereto.

[0056] The threshold value of palmitoleic acid is preferably 20 μEQ / L or more and 40 μEQ / L or less, more preferably 23 μEQ / L or more and 35 μEQ / L or less, and further preferably 25 μEQ / L or more and 33 μEQ / L or less. When the gynecological cancer is an ovarian malignancy, 29.06 μEQ / L can be cited as a specific example of the threshold value of palmitoleic acid. When early ovarian malignancy is to be detected, the same threshold value as above can also be cited as a specific example of the threshold value. When the gynecological cancer is cervical cancer, 29.12 μEQ / L can be cited as a specific example of the threshold value of palmitoleic acid. When precancerous lesions including cervical cancer are to be detected, 36.63 μEQ / L can be cited as a specific example of the threshold value of palmitoleic acid.

[0057] The threshold value of linoleic acid is preferably 170 μEQ / L or more and 200 μEQ / L or less, more preferably 175 μEQ / L or more and 190 μEQ / L or less, and further preferably 180 μEQ / L or more and 185 μEQ / L or less. When the gynecological cancer is an ovarian malignancy, 183.1 μEQ / L is a specific example of the threshold value of linoleic acid. When early ovarian malignancy is to be detected, the same threshold values ​​as above can also be cited as specific examples of the threshold value. When the gynecological cancer is cervical cancer, 84.63 μEQ / L is a specific example of the threshold value of linoleic acid. When precancerous lesions including cervical cancer are to be detected, 83.34 μEQ / L is a specific example of the threshold value of linoleic acid.

[0058] The threshold value of α-linolenic acid is preferably 20 μEQ / L or more and 40 μEQ / L or less, more preferably 23 μEQ / L or more and 35 μEQ / L or less, and further preferably 25 μEQ / L or more and 33 μEQ / L or less. When the gynecological cancer is an ovarian malignancy, 28.62 μEQ / L is a specific example of the threshold value of α-linolenic acid. When early ovarian malignancy is to be detected, the same threshold values ​​as above can also be cited as specific examples of the threshold value. When the gynecological cancer is cervical cancer, 13.13 μEQ / L is a specific example of the threshold value of α-linolenic acid. When precancerous lesions including cervical cancer are to be detected, 8.334 μEQ / L is a specific example of the threshold value of α-linolenic acid.

[0059] The threshold value of oleic acid is preferably 300 μEQ / L or more and 350 μEQ / L or less, more preferably 310 μEQ / L or more and 340 μEQ / L or less, and further preferably 320 μEQ / L or more and 330 μEQ / L or less. When the gynecological cancer is an ovarian malignancy, a specific example of the threshold value of oleic acid is 326.0 μEQ / L or less. When early ovarian malignancy is to be detected, the same threshold value as above can also be cited as a specific example of the threshold value. When the gynecological cancer is cervical cancer, a specific example of the threshold value of oleic acid is 251.5 μEQ / L. When precancerous lesions including cervical cancer are to be detected, a specific example of the threshold value of oleic acid is 333.4 μEQ / L.

[0060] The threshold value of vaccenic acid is preferably 1.2 μEQ / L or more and 2.4 μEQ / L or less, more preferably 1.5 μEQ / L or more and 2.2 μEQ / L or less, and further preferably 1.8 μEQ / L or more and 2.0 μEQ / L or less. When the gynecological cancer is an ovarian malignancy, 1.926 μEQ / L is a specific example of the threshold value of vaccenic acid. When early ovarian malignancy is to be detected, 1.930 μEQ / L is a specific example of the threshold value of vaccenic acid. When the gynecological cancer is cervical cancer, 5.69 μEQ / L is a specific example of the threshold value of vaccenic acid. When precancerous lesions including cervical cancer are to be detected, 6.655 μEQ / L is a specific example of the threshold value of arachidonic acid.

[0061] The threshold value of arachidic acid is preferably 1.2 μEQ / L or more and 2.4 μEQ / L or less, more preferably 1.5 μEQ / L or more and 2.2 μEQ / L or less, and still more preferably 1.8 μEQ / L or more and 2.0 μEQ / L or less. When the gynecological cancer is an ovarian malignancy, 1.924 μEQ / L is a specific example of the threshold value of arachidic acid. When early ovarian malignancy is to be detected, 1.953 μEQ / L is a specific example of the threshold value of arachidic acid. When the gynecological cancer is cervical cancer, 1.875 μEQ / L is a specific example of the threshold value of arachidic acid. When precancerous lesions including cervical cancer are to be detected, 1.781 μEQ / L is a specific example of the threshold value of arachidic acid.

[0062] The threshold value of docosahexaenoic acid is preferably 1.3 μEQ / L or more and 2.3 μEQ / L or less, more preferably 1.5 μEQ / L or more and 2.2 μEQ / L or less, and further preferably 1.7 μEQ / L or more and 2.03 μEQ / L or less. When the gynecological cancer is an ovarian malignant tumor, a specific example of the threshold value of docosahexaenoic acid is 1.847 μEQ / L. When early ovarian malignant tumors are to be detected, the same threshold values ​​as above can also be cited as specific examples of the threshold value. When the gynecological cancer is cervical cancer, a specific example of the threshold value of docosahexaenoic acid is 2.670 μEQ / L.

[0063] The threshold value of adrenal acid is preferably 0.55 μEQ / L or more and 0.85 μEQ / L or less, more preferably 0.60 μEQ / L or more and 0.80 μEQ / L or less, and further preferably 0.65 μEQ / L or more and 0.75 μEQ / L or less. When the gynecological cancer is an ovarian malignancy, a specific example of the threshold value of adrenal acid is 0.7085 μEQ / L. When the gynecological cancer is cervical cancer, a specific example of the threshold value of adrenal acid is 0.6688 μEQ / L.

[0064] The threshold value of neuraminic acid is preferably 0.06 μEQ / L or more and 0.11 μEQ / L or less, more preferably 0.07 μEQ / L or more and 0.10 μEQ / L or less, and further preferably 0.08 μEQ / L or more and 0.09 μEQ / L or less. When the gynecological cancer is an ovarian malignant tumor, a specific example of the threshold value of neuraminic acid is 0.0891 μEQ / L.

[0065] The threshold value of lignoceric acid is preferably 0.3 μEQ / L or more and 0.8 μEQ / L or less, more preferably 0.4 μEQ / L or more and 0.7 μEQ / L or less, and further preferably 0.5 μEQ / L or more and 0.6 μEQ / L or less. When the gynecological cancer is an ovarian malignant tumor, a specific example of the threshold value of lignoceric acid is 0.5773 μEQ / L. When early ovarian malignant tumors are to be detected, the same threshold values ​​as above can also be cited as specific examples of the threshold value.

[0066] The threshold value of the stearic acid concentration is preferably 200 μEQ / L to 300 μEQ / L, more preferably 220 μEQ / L to 280 μEQ / L, and further preferably 240 μEQ / L to 260 μEQ / L. Stearic acid can be suitably used for evaluation when the gynecological cancer is, for example, uterine cancer or cervical cancer.

[0067] As the threshold value of palmitic acid, preferably 350 μEQ / L or more and 600 μEQ / L or less, more preferably 400 μEQ / L or more and 550 μEQ / L or less, further preferably 430 μEQ / L or more and 500 μEQ / L or less. In the case where the gynecological cancer is ovarian cancer, a specific example of the threshold value of palmitic acid is 452.9 EQ / L. Palmitic acid can be suitably used for evaluation, for example, in the case where the gynecological cancer is uterine cancer or cervical cancer.

[0068] The threshold value of dihomo-γ-linolenic acid is preferably 0.8 μEQ / L or more and 1.0 μEQ / L or less, more preferably 0.85 μEQ / L or more and 0.95 μEQ / L or less, and further preferably 0.90 μEQ / L or more and 0.95 μEQ / L or less. When the gynecological cancer is cervical cancer, a specific example of the threshold value of dihomo-γ-linolenic acid is 0.9315 μEQ / L.

[0069] The threshold value of docosapentaenoic acid is preferably 0.6 μEQ / L or more and 1.5 μEQ / L or less, more preferably 0.65 μEQ / L or more and 1.2 μEQ / L or less, and still more preferably 0.7 μEQ / L or more and 1.0 μEQ / L or less. When the gynecological cancer is cervical cancer, a specific example of the threshold value of docosapentaenoic acid is 0.7814 μEQ / L.

[0070] The "predetermined threshold value" in the above (iii1) is a value set as the concentration ratio of docosapentaenoic acid to oleic acid. When the concentration ratio of docosapentaenoic acid to oleic acid calculated based on the free fatty acid concentration measured in step (A1) is lower than the threshold value set for the concentration ratio of docosapentaenoic acid to oleic acid, it is evaluated that the subject is likely to have one or more selected from the group consisting of gynecological cancer and precancerous lesions thereof.

[0071] The predetermined threshold value for the concentration ratio in (iii1) above can be a critical value obtained by calculating the concentration ratio from multiple samples from healthy female donors and multiple samples from patients with one or more types of cancer selected from a group consisting of gynecological cancer and its precancerous lesions, and performing statistical processing on these concentration ratios.

[0072] The predetermined threshold value for the concentration ratio in (iii1) above can also be set, for example, so that the positive likelihood ratio for the concentration ratio becomes maximum. The threshold value for the maximum positive likelihood ratio can be set, for example, by performing ROC analysis. For example, for a group of patients with one or more selected from a group consisting of gynecological cancer and precancerous lesions thereof, and a group of healthy female donors, their concentration ratios can be measured and ROC analysis can be performed using these concentration ratios.

[0073] When the sample derived from a subject used in the step (A1) is serum, an example of the predetermined threshold value for the concentration ratio of docosapentaenoic acid to oleic acid is 0.004433, but the present invention is not limited thereto.

[0074] In the step (B1), when the above (iii1) is met, it is preferably evaluated that the subject has a high possibility of cervical cancer. In this case, the detectable cervical cancer may be cervical cancer with a progression degree of stage I or higher.

[0075] When the detection object is gynecological cancer, the free fatty acid for evaluation is preferably selected from more than one free fatty acid in the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenal acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid. By selecting the free fatty acid for evaluation from the 14 kinds of free fatty acids, the detection accuracy of gynecological cancer is further improved. The free fatty acid for evaluation can be more than 2 kinds, more than 3 kinds, more than 4 kinds, more than 5 kinds, more than 6 kinds, more than 7 kinds, more than 8 kinds, more than 9 kinds, more than 10 kinds, more than 11 kinds, more than 12 kinds, more than 13 kinds or 14 kinds of free fatty acids in the free fatty acids. The free fatty acid for evaluation can be selected arbitrarily from these 14 kinds of fatty acids. When the detection target is gynecological cancer, for example, in step (B1), the following evaluation can be performed. If at least one of the following selected from the group consisting of (i1)-(0) and (ii1)-(0) is satisfied, it is evaluated that the subject is highly likely to have ovarian cancer; (i1)-0 The concentration of at least one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid and palmitic acid in the sample is higher than a predetermined threshold value; (ii1)-0 The concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid in the sample is lower than a predetermined threshold value.

[0076] In the case where the gynecological cancer that can be used as the detection object is ovarian cancer, the free fatty acid used for evaluation is preferably selected from one or more free fatty acids in the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid and lignoceric acid. By selecting the free fatty acid for evaluation from the 10 free fatty acids, the detection accuracy of ovarian cancer is further improved. The free fatty acid used for evaluation can be 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more or 10 free fatty acids in the free fatty acids. The free fatty acid used for evaluation can be arbitrarily selected from these 10 fatty acids. When the gynecological cancer is ovarian cancer, for example, the following evaluation can be performed in step (B1). If at least one of the following selected from the group consisting of (i1)-1 and (ii1)-1 is met, it is evaluated that the subject is highly likely to have ovarian cancer; (i1)-1 The concentration of at least one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid and oleic acid in the sample is higher than a predetermined threshold value; (ii1)-1 The concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid and lignoceric acid in the sample is lower than a predetermined threshold value.

[0077] In order to improve the detection sensitivity of early gynecological malignancies (e.g., stages I and II), the free fatty acids for evaluation can be free fatty acids selected from more than one of the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, and lignoceric acid. By selecting the free fatty acids for evaluation from the 8 free fatty acids, the detection accuracy of early gynecological malignancies (particularly ovarian malignancies of stage I or II) is further improved. The free fatty acids for evaluation can be more than 2, more than 3, more than 4, more than 5, more than 6, more than 7, or 8 free fatty acids in the free fatty acids. The free fatty acids for evaluation can be selected arbitrarily from these 8 fatty acids. When early-stage gynecological malignancies are to be detected, for example, the following evaluation may be performed in step (B1). In the case of at least one condition selected from the group consisting of the following (i1)-2 and (ii1)-2, the subject is evaluated to have a high possibility of having a gynecological cancer (e.g., ovarian malignant tumor) with a degree of progression higher than stage I; (i1)-2 The concentration of at least one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid and oleic acid in the sample is higher than a predetermined threshold value; (ii1)-2 The concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, arachidic acid, docosahexaenoic acid and lignoceric acid in the sample is lower than a predetermined threshold value.

[0078] In the case where the gynecological cancer that can be used as the detection object is uterine body cancer or cervical cancer, the free fatty acid for evaluation is preferably selected from more than one free fatty acid in the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenal acid and docosapentaenoic acid. By selecting the free fatty acid for evaluation from the 12 kinds of free fatty acids, the detection accuracy of uterine body cancer or cervical cancer is further improved. The free fatty acid for evaluation can be more than 2 kinds, more than 3 kinds, more than 4 kinds, more than 5 kinds, more than 6 kinds, more than 7 kinds, more than 8 kinds, more than 9 kinds, more than 10 kinds, more than 11 kinds or 12 kinds of free fatty acids in the free fatty acids. The free fatty acid for evaluation can be arbitrarily selected from these 12 kinds of fatty acids. When the gynecological cancer is uterine cancer or cervical cancer, for example, the following evaluation may be performed in step (B1). If at least one of the following selected from the group consisting of (i1)-3 and (ii1)-3 is met, it is evaluated that the subject is highly likely to have ovarian cancer; (i1)-3 the concentration of at least one free fatty acid selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid and oleic acid in the sample is higher than a predetermined threshold value; (ii1)-3 The concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid in the sample is lower than a predetermined threshold value.

[0079] In the case where the gynecological cancer that can be used as the detection object is uterine cancer, the free fatty acid for evaluation is preferably selected from one or more free fatty acids in the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid. By selecting the free fatty acid for evaluation from the 8 kinds of free fatty acids, the detection accuracy of uterine cancer is further improved. The free fatty acid for evaluation can be more than 2 kinds, more than 3 kinds, more than 4 kinds, more than 5 kinds, more than 6 kinds, more than 7 kinds or 8 kinds of free fatty acids in the free fatty acids. The free fatty acid for evaluation can be arbitrarily selected from these 8 kinds of fatty acids. When the gynecological cancer is uterine cancer, for example, the following evaluation may be performed in step (B1). If at least one of the following (i1)-4 and (ii1)-4 is met, it is evaluated that the subject is highly likely to have ovarian cancer; (i1)-4 The concentration of at least one free fatty acid selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid and oleic acid in the sample is higher than a predetermined threshold value; (ii1)-4 The concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, arachidic acid, docosahexaenoic acid and adrenic acid in the sample is lower than a predetermined threshold value.

[0080] In the case where the gynecological cancer that can be used as the detection object is cervical cancer, the free fatty acid used for evaluation is preferably selected from one or more free fatty acids in the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid. By selecting the free fatty acid used for evaluation from the 12 free fatty acids, the detection accuracy of cervical cancer is further improved. The free fatty acid used for evaluation can be 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more or 12 free fatty acids in the free fatty acids. The free fatty acid used for evaluation can be arbitrarily selected from these 12 fatty acids. When the gynecological cancer is cervical cancer, for example, the following evaluation may be performed in step (B1). If at least one condition selected from the group consisting of the following (i1) to (-5) and (ii1) to (-5) is satisfied, it is evaluated that the subject is highly likely to have ovarian cancer; (i1)-5 The concentration of at least one free fatty acid selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid and oleic acid in the sample is higher than a predetermined threshold value; (ii1)-5 The concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid in the sample is lower than a predetermined threshold value.

[0081] In the case of detecting gynecological cancer precancerous lesions (e.g., cervical cancer precancerous lesions), the free fatty acids used for evaluation are preferably free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, arachidonic acid and arachidic acid. The free fatty acids used for evaluation can be 2 or more, 3 or more, 4 or more, 5 or more, or 6 free fatty acids in the free fatty acids. The free fatty acids used for evaluation can be arbitrarily selected from these 6 fatty acids. When the detection target is gynecological cancer and its precancerous lesion, for example, in step (B1), the following evaluation may be performed. If at least one of the following (i1)-6 and (ii1)-6 is satisfied, the subject is evaluated to have a high possibility of having a gynecological cancer precancerous lesion and / or a gynecological cancer; (i1)-6 The concentration of at least one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid and oleic acid in the sample is higher than a predetermined threshold value; (ii1) The concentration of free fatty acid of -6-arachidonic acid in the sample is lower than a predetermined threshold value.

[0082] In step (B1), the step of determining whether or not one or more selected from the group consisting of the above-mentioned (i1) and (ii1) is met may be a step of determining that the object is likely to have gynecological cancer when the total value of the scores of the free fatty acids set in the following (i-1) and (ii-1) (hereinafter referred to as "FFA score") is higher than a predetermined threshold value (hereinafter also referred to as "step (B1-1)").

[0083] (i-1) When the free fatty acid is a free fatty acid of group B(i1), if the concentration of the free fatty acid in the sample is higher than a predetermined threshold, the score of the free fatty acid is set to "+1". If the concentration of the free fatty acid in the sample is lower than a predetermined threshold, the score of the free fatty acid is set to "0". (ii-1) When the free fatty acid is a free fatty acid of group B(ii1), if the concentration of the free fatty acid in the sample is lower than a predetermined threshold, the score of the free fatty acid is set to "+1". If the concentration of the free fatty acid in the sample is higher than a predetermined threshold, the score of the free fatty acid is set to "0". As the predetermined threshold value for each free fatty acid used in (i-1) and (ii-1), the same threshold value as the threshold value listed as the predetermined threshold value in (i1) and (ii1) above can be cited. The total value of the score of each free fatty acid set by (i-1) and (ii-1) is calculated as the FFA score.

[0084] The predetermined threshold value for the FFA score may also be a critical value obtained by calculating FFA scores for multiple samples from healthy female donors and multiple samples from patients with one or more selected from the group consisting of gynecological cancer and precancerous lesions thereof, and performing statistical processing on these FFA scores.

[0085] The predetermined threshold value for the FFA score can also be set, for example, so that the positive likelihood ratio for the FFA score becomes the maximum. The threshold value for which the positive likelihood ratio becomes the maximum can be set, for example, by performing ROC analysis. For example, for an arbitrary number of patients having one or more selected from a group consisting of gynecological cancer and precancerous lesions thereof, and a female healthy donor group, the serum concentration of a specific free fatty acid can be measured, and these serum concentrations can be used to perform ROC analysis.

[0086] The predetermined threshold value for the FFA score also depends on the number of free fatty acids used in the calculation of the FFA score, and for example, 1 or more and 3 or less, preferably greater than 1 and less than 3, more preferably greater than 1 and less than 2, and further preferably greater than 1 and less than 2. As the number of free fatty acids used in the calculation of the FFA score, for example, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, and 15 of the free fatty acids in the above-mentioned A1 group can be cited. The free fatty acids used in the calculation of the FFA score can be arbitrarily selected from the fatty acids in the A1 group.

[0087] In the case where gynecological cancer is ovarian cancer, as the free fatty acid for calculating the FFA score, it is preferably selected from more than one of the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenal acid, nervonic acid and lignoceric acid. The free fatty acid used in the calculation of the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9 or 10 of the free fatty acid. The free fatty acid for calculating the FFA score can be arbitrarily selected from these fatty acids. In the case where gynecological cancer is ovarian cancer, in the calculation of the FFA score, it is preferred to use the 10 kinds of free fatty acids. When using the 10 kinds of free fatty acids, as the predetermined threshold value of the FFA score, the same threshold value as above can be cited, for example, it can be set to be greater than 1 and less than 2 (for example 1.5).

[0088] In order to improve the detection sensitivity of early gynecological malignancies (such as stage I, II), the free fatty acids used to calculate the FFA score can be one or more selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid and lignoceric acid. The free fatty acids used to calculate the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7 or 8 of the free fatty acids. The free fatty acids used to calculate the FFA score can be arbitrarily selected from these fatty acids. From the viewpoint of the detection sensitivity of early gynecological malignancies, the calculation of the FFA score preferably uses the 8 free fatty acids. When using the 8 free fatty acids, as the predetermined threshold value of the FFA score, the same threshold value as above can be cited, for example, it can be set to be more than 1 and less than 2 (for example 1.5).

[0089] In the case where gynecological cancer is uterine body cancer or cervical cancer, as the free fatty acid for calculating the FFA score, it is preferably selected from more than one of the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenal acid and docosapentaenoic acid. The free fatty acid for calculating the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9, more than 10, more than 11 or 12 kinds of the free fatty acid. The free fatty acid for calculating the FFA score can be arbitrarily selected from these fatty acids. The number of species of the free fatty acids for calculating the FFA score and the predetermined threshold value of the FFA score can be appropriately set.

[0090] In the case where gynecological cancer is uterine cancer, as the free fatty acid for calculating the FFA score, preferably one or more selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid. The free fatty acid for calculating the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7 or 8 of the free fatty acids. The free fatty acid for calculating the FFA score can be arbitrarily selected from these fatty acids. The number of species of the free fatty acids for calculating the FFA score and the predetermined threshold value of the FFA score can be appropriately set.

[0091] In the case where gynecological cancer is cervical cancer, as the free fatty acid for calculating the FFA score, it is preferably selected from more than one of the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid. The free fatty acid for calculating the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9, more than 10, more than 11 or 12 kinds of the free fatty acid. The free fatty acid for calculating the FFA score can be arbitrarily selected from these fatty acids. The number of species of the free fatty acids for calculating the FFA score and the predetermined threshold value of the FFA score can be appropriately set.

[0092] In the case of detecting gynecological cancer precancerous lesions, as the free fatty acid for calculating the FFA score, it is preferably selected from one or more of the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, arachidonic acid and arachidic acid. The free fatty acid for calculating the FFA score can be 2 or more, 3 or more, 4 or more, 5 or more or 6 of the free fatty acids. The free fatty acid for calculating the FFA score can be arbitrarily selected from these fatty acids. The number of types of free fatty acids for calculating the FFA score and the predetermined threshold value of the FFA score can be appropriately set.

[0093] In the inspection method of the first embodiment, when the FFA score is used, specific examples of the step (A1) and the step (B1) include the following. (Example 1) In step (A1), the concentrations of 10 free fatty acids consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, and lignoceric acid in a sample derived from a subject are measured. In step (B1), when the FFA score calculated based on 10 free fatty acids consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid and lignoceric acid is higher than a predetermined threshold, it is determined that the subject is likely to have gynecological cancer. The threshold value of the FFA score calculated from the above 10 free fatty acids is preferably 1 to 3, more preferably 1.2 to 2.5, further preferably 1.3 to 2, particularly preferably 1.3 to 1.9.

[0094] (Example 2) In step (A1), the concentrations of eight free fatty acids consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, and lignoceric acid in a sample derived from a subject are measured. In step (B1), when the FFA score calculated based on eight free fatty acids consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid and lignoceric acid is higher than a predetermined threshold, it is determined that the subject is likely to have gynecological cancer. The threshold value of the FFA score of the eight free fatty acids is preferably 0.5 or more and 2.5 or less, more preferably 1 or more and 2 or less, and even more preferably 1.1 or more and 1.9 or less.

[0095] By using FFA scoring, it is possible to more accurately evaluate the possibility that an object has one or more selected from a group consisting of gynecological cancer and its precancerous lesions. By using FFA scoring, even early gynecological malignancies (e.g., early ovarian malignancies), gynecological clear cell malignancies (e.g., ovarian clear cell malignancies), or gynecological borderline malignancies (e.g., ovarian borderline malignancies) can be detected with high accuracy. These tumors have a high proportion of false negatives in previous tumor markers such as CA125, and low sensitivity. However, by using FFA scoring, these tumors can also be detected with high sensitivity. For example, regarding the early gynecological malignancies, by using FFA scoring, gynecological malignancies in stage I or stage II can also be detected with high sensitivity.

[0096] In step (B1), the subject evaluated as having a high probability of having one or more selected from the group consisting of gynecological cancer and its precancerous lesions has a high probability of having one or more selected from the group consisting of any one of the gynecological cancers and its precancerous lesions. Therefore, in order to determine the type of one or more selected from the group consisting of gynecological cancer and its precancerous lesions that the subject has, further examinations may be performed by computed tomography (CT), magnetic resonance imaging (MRI), positron emission tomography (PET), ultrasonic examination, etc. Furthermore, for a confirmed diagnosis, a pathological examination (cytodiagnostic examination, histological examination) may also be performed.

[0097] The sample inspection method of the first aspect may further include the following step (C1). Step (C1): For the subject determined by the step (B1) to have a high probability of having one or more selected from the group consisting of gynecological cancer and its precancerous lesions, evaluating which of gynecological cancer and gynecological cancer precancerous lesions the subject has based on the concentration of the free fatty acid obtained in the step (A1) in the sample.

[0098] The subject in step (C1) is a subject determined in step (B1) to have a high possibility of having one or more selected from the group consisting of gynecological cancer and precancerous lesions thereof.

[0099] (Process (C1)) In step (B1), based on the concentration of the free fatty acid in the sample obtained in step (A1), it is determined whether the subject has gynecological cancer or gynecological cancer precancerous lesion.

[0100] The gynecological cancer to be determined is preferably cervical cancer. The gynecological cancer precancerous lesion to be determined is preferably cervical cancer precancerous lesion.

[0101] In step (C1), when the following (Ci) is satisfied, it is evaluated that the subject is highly likely to have gynecological cancer. (Ci) The concentration of at least one free fatty acid selected from the group consisting of dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid in the sample is lower than a predetermined threshold value.

[0102] Hereinafter, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid are also collectively referred to as "free fatty acids of group C1".

[0103] In step (C1), the number of types of free fatty acids to be determined may be 1 or more, 2 or more, 3 or more, or 4, as long as it is selected from the free fatty acids in group C1. The free fatty acids to be determined may be arbitrarily selected from the free fatty acids in group C1.

[0104] The "predetermined threshold value" in the above (Ci) is a value set for each free fatty acid. The predetermined threshold value of the concentration of each free fatty acid in the above (Ci) can be a critical value obtained by obtaining the concentration of each free fatty acid from multiple samples from patients with gynecological cancer and multiple samples from patients with gynecological cancer precancerous lesions, and performing statistical processing on the concentrations of each free fatty acid.

[0105] The predetermined threshold value of the concentration of a specific free fatty acid can be set, for example, so that the positive likelihood ratio for the specific free fatty acid becomes the maximum. The threshold value at which the positive likelihood ratio becomes the maximum can be set, for example, by performing ROC analysis. For example, for a patient group with gynecological cancer and a patient group with gynecological cancer precancerous lesions of any number of people, the serum concentration of a specific free fatty acid can be measured, and these serum concentrations are used to perform ROC analysis.

[0106] Examples of the threshold value of the concentration of each free fatty acid in (Ci) include the following threshold values, but the present invention is not limited thereto.

[0107] The threshold value of dihomo-γ-linolenic acid is preferably 0.80 μEQ / L or more and 1.20 μEQ / L or less, more preferably 0.90 μEQ / L or more and 1.10 μEQ / L or less, and further preferably 0.95 μEQ / L or more and 1.05 μEQ / L or less. When the gynecological cancer is cervical cancer and the gynecological cancer precancerous lesion is a cervical cancer precancerous lesion, a specific example of the threshold value of dihomo-γ-linolenic acid is 1.012 μEQ / L.

[0108] The threshold value of docosahexaenoic acid is preferably 2.0 μEQ / L or more and 5.0 μEQ / L or less, more preferably 2.5 μEQ / L or more and 4.5 μEQ / L or less, and still more preferably 3.0 μEQ / L or more and 4.0 μEQ / L or less. When the gynecological cancer is cervical cancer and the gynecological cancer precancerous lesion is a cervical cancer precancerous lesion, a specific example of the threshold value of docosahexaenoic acid is 3.507 μEQ / L.

[0109] The threshold value of adrenaline is preferably 0.15 μEQ / L or more and 0.60 μEQ / L or less, more preferably 0.20 μEQ / L or more and 0.50 μEQ / L or less, and still more preferably 0.25 μEQ / L or more and 0.40 μEQ / L or less. When the gynecological cancer is cervical cancer and the gynecological cancer precancerous lesion is a cervical cancer precancerous lesion, a specific example of the threshold value of docosahexaenoic acid is 0.312 μEQ / L.

[0110] The threshold value of docosapentaenoic acid is preferably 0.40 μEQ / L or more and 1.00 μEQ / L or less, more preferably 0.50 μEQ / L or more and 0.90 μEQ / L or less, and still more preferably 0.60 μEQ / L or more and 0.80 μEQ / L or less. When the gynecological cancer is cervical cancer and the gynecological cancer precancerous lesion is a cervical cancer precancerous lesion, a specific example of the threshold value of docosapentaenoic acid is 0.7231 μEQ / L.

[0111] Step (C1) may also be a step of calculating an FFA score, and evaluating that the subject has a high possibility of having gynecological cancer when the FFA score is higher than a predetermined threshold value. The FFA score is calculated by the same method as described in the first embodiment. That is, the FFA score is calculated as the total value of the scores of each free fatty acid set in the same manner as in (i-1) and (ii-1) above.

[0112] The predetermined threshold value for the FFA score may be a critical value obtained by calculating the FFA scores for a plurality of samples from gynecological cancer patients and a plurality of samples from patients with precancerous lesions and performing statistical processing on the FFA scores.

[0113] The predetermined threshold value for the FFA score also depends on the number of free fatty acid species used in the calculation of the FFA score. For example, when the number of free fatty acid species is set to n, examples include 0.2n or more and 0.6n or less, 0.3n or more and 0.6n or less, or 0.4n or more and 0.6n or less. As the number of free fatty acid species used to calculate the FFA score, for example, three or more free fatty acids in the above-mentioned A5 group or four free fatty acids can be mentioned.

[0114] Through the diagnosis as described above, for an object confirmed to have one or more selected from the group consisting of gynecological cancers and precancerous lesions, one or more treatments selected from the group consisting of gynecological cancers and precancerous lesions can be performed. In one embodiment, the present invention provides a method of treating one or more selected from the group consisting of gynecological cancers and precancerous lesions, comprising: (a1) a process of determining an object with a high probability of having one or more selected from the group consisting of gynecological cancers and precancerous lesions by a first method of sample inspection; (b1) a process of confirming that the object has one or more selected from the group consisting of gynecological cancers and precancerous lesions; and (c1) a process of performing one or more treatments selected from the group consisting of gynecological cancers and precancerous lesions on the object confirmed to have one or more selected from the group consisting of gynecological cancers and precancerous lesions. In the step (a1), as described above, the sample testing method of the first aspect is carried out to identify a subject with a high possibility of having one or more selected from the group consisting of gynecological cancer and precancerous lesions thereof. In the step (b1), by further testing the subject identified in the step (a1), it is possible to confirm that the subject has one or more selected from the group consisting of gynecological cancer and precancerous lesions thereof. As one or more tests selected from the group consisting of gynecological cancer and precancerous lesions thereof, for example, the above-mentioned tests can be cited. In the step (c1), for an object confirmed in the step (b1) to have one or more selected from the group consisting of gynecological cancer and its precancerous lesions, one or more treatments selected from the group consisting of gynecological cancer and its precancerous lesions can be implemented. As a treatment method for gynecological cancer, there is no particular limitation, and for example, surgical treatment, chemotherapy, and radiotherapy can be cited. For example, in the case of early gynecological cancer (for example, stage I or stage II), surgical treatment can be selected as the main treatment method. In the case of early gynecological cancer, for example, if the gynecological cancer can be completely removed by surgical treatment, the gynecological cancer can be completely cured. In the case of gynecological cancer with advanced progression (for example, stage III or stage IV), surgical treatment, chemotherapy (administration of anticancer agents, etc.) and radiotherapy can also be combined for treatment. For example, chemotherapy and / or radiotherapy can be performed before surgical treatment to cause tumor regression. Alternatively, chemotherapy and / or radiotherapy can also be performed after surgical treatment to cause residual tumors to regress after surgical treatment. Examples of anticancer agents used in chemotherapy include the anticancer agents described later in the section [Medical Products]. As treatment methods for gynecological cancer precancerous lesions, vaginal surgeries such as cervical conization and cervical laser ablation can be performed.

[0115] According to the sample inspection method of the first aspect described above, it is possible to easily determine the possibility that a subject has one or more selected from the group consisting of gynecological cancer and precancerous lesions thereof with high specificity and high sensitivity. Diagnosis using CA125 as a marker has low sensitivity for detecting early ovarian malignancies and ovarian clear cell malignancies. The first aspect of the inspection method can detect early ovarian malignancies or ovarian clear cell malignancies with high specificity and sensitivity even when the subject has early ovarian malignancies or ovarian clear cell malignancies.

[0116] Even if treatment is given to a gynecological cancer that is progressing, the possibility of recurrence is high, and there are many cases where the prognosis is poor. On the other hand, when gynecological cancer is detected at an early stage before progression, treatment such as tumor resection can completely cure the gynecological cancer. Alternatively, the prognosis can be improved. According to the inspection method of the sample involved in the first embodiment, gynecological cancer can be detected at an early stage, so the probability of curing gynecological cancer through treatment can be increased.

[0117] Since cervical cancer has the risk of metastasis, highly invasive surgical treatments such as quasi-extensive / extensive total hysterectomy and lymph node dissection or treatments based on radiation exposure are performed. Therefore, fertility is sometimes cut off in young people aged 30-40, who are at the peak of the disease. On the other hand, cervical precancerous lesions are tumors confined to the epithelium without infiltration, so there is no possibility of metastasis. Therefore, low-invasive treatments such as cervical cone resection and cervical laser ablation are performed, and fertility is also preserved. Therefore, detecting cervical precancerous lesions becomes a great advantage for the subject.

[0118] Step (B1) may be a step of determining a sample derived from a subject with a high probability of having one or more selected from the group consisting of gynecological cancer and its precancerous lesions when at least one selected from the group consisting of (i1) and (ii1) is satisfied.

[0119] Step (C1) may be a step of specifying a sample derived from a subject with a high possibility of having the gynecological cancer when the condition (Ci) is met.

[0120] [Test sample inspection method (2)] The second aspect of the sample testing method is a method for evaluating the possibility of a poor prognosis in a subject with gynecological cancer. The testing method includes step (A2) and step (B2). Step (A2): a step of measuring the concentration of one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid in a sample derived from a subject Step (B2): a step of evaluating the possibility of poor prognosis based on the concentration of free fatty acids in the sample obtained in step (A2)

[0121] In the second method of testing a sample, the sample for testing is derived from a human female with gynecological cancer. For example, the subject is a human female who has been pre-diagnosed as having gynecological cancer. The subject may be before the start of treatment for gynecological cancer, or may be in the process of treatment for gynecological cancer, or may have received treatment for gynecological cancer in the past. Alternatively, the subject may be determined to have gynecological cancer by the first method of testing. Examples of the gynecological cancer include the cancers described in the first aspect. The gynecological cancer is preferably a gynecological malignant tumor, more preferably one or more selected from the group consisting of an ovarian malignant tumor, a uterine body malignant tumor, and a cervical malignant tumor, and even more preferably an ovarian malignant tumor.

[0122] Palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid are also collectively referred to as "free fatty acids of group A2".

[0123] Step (A2) can be performed in the same manner as step (A1) in the first embodiment, except that a sample derived from a subject having gynecological cancer is used. The measurement may be performed in vitro.

[0124] In step (A2), the number of types of free fatty acids to be measured may be one or more of the free fatty acids selected from group A2, may be two or more, may be three or more, may be four or more, may be five or more, may be six or more, may be seven or more, may be eight or more, may be nine or more, may be ten or more, may be eleven or more, may be twelve or more, may be thirteen or more, may be fourteen. The free fatty acids to be measured may be arbitrarily selected from the free fatty acids in group A2.

[0125] When the gynecological cancer is ovarian cancer, in step (A2), one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid and lignoceric acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these free fatty acids. When the gynecological cancer is ovarian cancer, in step (A2), the concentrations of the ten free fatty acids are preferably measured.

[0126] When gynecological cancer is an early gynecological malignancy (e.g., stage I, II), in step (A2), one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, and tetracosanoic acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, or eight free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these free fatty acids. When early gynecological malignancies are to be detected, the concentrations of the eight free fatty acids are preferably measured in step (A2). As early gynecological malignancies, for example, early ovarian malignancies can be cited.

[0127] When gynecological cancer is uterine corpus cancer or cervical cancer, in step (A2), one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenal acid and docosapentaenoic acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, or twelve free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these free fatty acids. When gynecological cancer is uterine corpus cancer or cervical cancer, in step (A2), the concentrations of the twelve free fatty acids are preferably measured.

[0128] When gynecological cancer is uterine cancer, in step (A2), one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, or eight free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these free fatty acids. When gynecological cancer is uterine cancer, in step (A2), the concentrations of the eight free fatty acids are preferably measured.

[0129] When the gynecological cancer is cervical cancer, in step (A2), one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, or twelve free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these free fatty acids. When the detection object is cervical cancer, in step (A2), the concentrations of the twelve free fatty acids are preferably measured.

[0130] In step (B2), when at least one selected from the group consisting of the following (i2) and (ii2) is satisfied, it is evaluated that the subject has a high possibility of having a poor prognosis. (i2) the concentration of at least one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid and palmitic acid in the sample is higher than a predetermined threshold value; (ii2) the concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid in the sample is lower than a predetermined threshold value.

[0131] In the case where the gynecological cancer is ovarian cancer, the free fatty acid used in the evaluation is preferably selected from one or more of the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid and lignoceric acid. The free fatty acid used for evaluation can be 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more or 10 of the free fatty acids. The free fatty acid used for evaluation can be arbitrarily selected from the 10 free fatty acids. In the case where the gynecological cancer is ovarian cancer, the free fatty acid used for evaluation is preferably the 10 free fatty acids.

[0132] In the case where gynecological cancer is an early gynecological malignancy, the free fatty acid used in the evaluation is preferably selected from one or more of the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid and tetracosanoic acid. The free fatty acid used for evaluation can be 3 or more, 4 or more, 5 or more, 6 or more, 7 or more or 8 of the free fatty acids. The free fatty acid used for evaluation can be arbitrarily selected from the 8 free fatty acids. In the case where an early gynecological malignancy is to be detected, the free fatty acid used for evaluation is preferably the 8 free fatty acids.

[0133] In the case where gynecological cancer is uterine corpus cancer or cervical cancer, as the free fatty acid used in the evaluation, it is preferably selected from more than one of the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenal acid and docosapentaenoic acid. The free fatty acid used for evaluation can be more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9, more than 10, more than 11 or 12 kinds of the free fatty acid. The free fatty acid used for evaluation can be arbitrarily selected from the 12 kinds of free fatty acids. In the case where gynecological cancer is uterine corpus cancer or cervical cancer, the free fatty acid used for evaluation is preferably the 12 kinds of free fatty acids.

[0134] In the case where gynecological cancer is uterine corpus cancer, as the free fatty acid used in the evaluation, it is preferably selected from more than one of the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid. The free fatty acid used for evaluation can be more than 3, more than 4, more than 5, more than 6, more than 7 or 8 of the free fatty acids. The free fatty acid used for evaluation can be arbitrarily selected from these free fatty acids. In the case where gynecological cancer is uterine corpus cancer, the free fatty acid used for evaluation is preferably the 12 kinds of free fatty acids.

[0135] In the case where the gynecological cancer is cervical cancer, the free fatty acid used in the evaluation is preferably selected from one or more of the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid. The free fatty acid used for evaluation can be 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more or 12 of the free fatty acids. The free fatty acid used for evaluation can be arbitrarily selected from these free fatty acids. In the case where the gynecological cancer is cervical cancer, the free fatty acid used for evaluation is preferably the 12 kinds of free fatty acids.

[0136] The "predetermined threshold value" in (i2) and (ii2) above is a value set for each free fatty acid. The predetermined threshold value of the concentration of each free fatty acid can be a critical value obtained by obtaining the concentration of each free fatty acid based on multiple samples from gynecological patients with good prognosis and multiple samples from gynecological cancer patients with poor prognosis, and performing statistical processing on the concentrations of each free fatty acid.

[0137] Step (B2) may also be a step of calculating an FFA score, and evaluating that the subject has a high probability of having a poor prognosis for gynecological cancer when the FFA score is higher than a predetermined threshold value. The FFA score is calculated by the same method as described in the first embodiment. That is, the FFA score is calculated as the total value of the scores of the free fatty acids set in the same manner as in (i-1) and (ii-1) above.

[0138] In step (B2), the predetermined threshold value set in the same manner as in (i-1) and (ii-1) above may be acquired as described above, or the same threshold value as in the inspection method of the first embodiment may be used. The predetermined threshold value for the FFA score may also be a critical value obtained by calculating the FFA scores for a plurality of samples from gynecological cancer patients with a good prognosis and a plurality of samples from gynecological cancer patients with a poor prognosis, and performing statistical processing on these FFA scores. Alternatively, the FFA scores may be calculated for a plurality of samples from gynecological cancer patients, and the median value of these FFA scores may be used as the threshold value for the FFA score.

[0139] The predetermined threshold value about FFA score also depends on the number of species of free fatty acids used in the calculation of FFA score, for example, when the number of species of free fatty acids is set to n, more than 0.4n and less than 0.8n, more than 0.5n and less than 0.7n, or more than 0.5n and less than 0.6n, etc. can be cited. As the number of species of free fatty acids used in the calculation of FFA score, for example, more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9 or more than 10 species in the free fatty acids of the above-mentioned A2 group can be cited. "0.4n" refers to the value obtained by multiplying 0.4 by the number of species n. It is the same for other numerical values. For example, when the number of species of free fatty acids is 5, "more than 0.4n and less than 0.8n" refers to "more than 2 and less than 4".

[0140] In the case where gynecological cancer is ovarian cancer, as the free fatty acid for calculating the FFA score, it is preferably selected from more than one of the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid and lignoceric acid. The free fatty acid for calculating the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9 or 10 of the free fatty acids. The free fatty acid for calculating the FFA score can be arbitrarily selected from these free fatty acids. In the case where gynecological cancer is ovarian cancer, in the calculation of the FFA score, it is preferred to use the 10 free fatty acids. The predetermined threshold value for the FFA score may be the same as the predetermined threshold value for FFA described in step (B2). When the 10 free fatty acids are used, the predetermined threshold value for the FFA score may be, for example, more than 4 and less than 6.

[0141] In the case where gynecological cancer is an early gynecological malignancy, the free fatty acid used in the calculation of the FFA score is preferably selected from more than one of the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid and lignoceric acid. The free fatty acid used to calculate the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7 or 8 of the free fatty acids. The free fatty acid used to calculate the FFA score can be arbitrarily selected from these free fatty acids. In the case of detecting early gynecological malignancies, it is preferred to use the 8 free fatty acids in the calculation of the FFA score. The predetermined threshold value for the FFA score may be the same as the predetermined threshold value for FFA described in step (B2). When the eight free fatty acids are used, the predetermined threshold value for the FFA score may be, for example, greater than 3 and less than 5.

[0142] In the case where gynecological cancer is uterine corpus cancer or cervical cancer, as the free fatty acid for calculating the FFA score, preferably one or more selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenal acid and docosapentaenoic acid. The free fatty acid used in the calculation of the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9, more than 10, more than 11 or 12 kinds of the free fatty acid. The free fatty acid for calculating the FFA score can be arbitrarily selected from these free fatty acids. The number of species of the free fatty acids for calculating the FFA score, the predetermined threshold value of the FFA score can be appropriately set. In the case where gynecological cancer is uterine corpus cancer or cervical cancer, in the calculation of the FFA score, the 12 kinds of free fatty acids are preferably used. The predetermined threshold value for the FFA score may be the same as the predetermined threshold value for FFA described in step (B2).

[0143] In the case where gynecological cancer is uterine corpus cancer, as the free fatty acid for calculating the FFA score, it is preferably selected from more than one of the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid. The free fatty acid for calculating the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7 or 8 of the free fatty acids. The free fatty acid for calculating the FFA score can be arbitrarily selected from these free fatty acids. The number of species of free fatty acids for calculating the FFA score and the predetermined threshold value of the FFA score can be appropriately set. In the case where gynecological cancer is uterine corpus cancer, the 8 kinds of free fatty acids are preferably used in the calculation of the FFA score. The predetermined threshold value for the FFA score may be the same as the predetermined threshold value for FFA described in step (B2).

[0144] In the case where gynecological cancer is cervical cancer, as the free fatty acid for calculating the FFA score, it is preferably selected from more than one of the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenal acid and docosapentaenoic acid. The free fatty acid for calculating the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9, more than 10, more than 11 or 12 kinds of the free fatty acid. The free fatty acid for calculating the FFA score can be arbitrarily selected from these free fatty acids. The number of species of free fatty acids for calculating the FFA score and the predetermined threshold value of the FFA score can be appropriately set. In the case where gynecological cancer is cervical cancer, the calculation of the FFA score preferably uses the 10 free fatty acids. The predetermined threshold value for the FFA score may be the same as the predetermined threshold value for FFA described in step (B2).

[0145] In the inspection method of the second embodiment, when the FFA score is used, specific examples of the step (A2) and the step (B2) include the following. In step (A2), the concentrations of 10 free fatty acids consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, and lignoceric acid in a sample derived from a subject are measured. In step (B2), when the FFA score of 10 free fatty acids consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid and lignoceric acid is higher than a predetermined threshold value, it is evaluated that the subject is likely to have a poor prognosis. The predetermined threshold value for the FFA score may also be a critical value obtained by calculating the FFA scores for 10 free fatty acids from gynecological cancer patients with good prognosis and 10 free fatty acids from gynecological cancer patients with poor prognosis, and performing statistical processing on these FFA scores. Alternatively, the FFA scores may be calculated for a plurality of samples from gynecological cancer patients, and the median value of these FFA scores may be used as the threshold value for the FFA score.

[0146] According to the inspection method of the sample involved in the second mode described above, it is possible to predict whether the object has a poor prognosis. Therefore, it is easy to formulate an appropriate treatment plan with reference to its inspection results. In addition, it is easy to predict recurrence after treatment. When the score is higher than a predetermined threshold value, it is evaluated that the object has a high possibility of having a poor prognosis.

[0147] For example, for a subject evaluated as having a high probability of having a poor prognosis, measures such as changing the treatment method may also be taken. In one embodiment, the present invention provides a treatment method for gynecological cancer, comprising: (a2) determining a subject having a high probability of having a poor prognosis by the sample inspection method involved in the second embodiment; and (b2) changing the treatment method for gynecological cancer for the subject. In the step (a2), the sample testing method according to the second aspect is carried out as described above to identify subjects with a high possibility of having a poor prognosis. In the step (b2), for the object determined in the step (a2), the treatment method of gynecological cancer can be studied again, and the treatment method of gynecological cancer can be changed. Even if the object determined in the step (a2) continues the previous treatment, the possibility of becoming a poor prognosis is high. Therefore, the possibility of improving the prognosis can be explored by studying the treatment method again and changing the treatment method. As a change in the treatment method, for example, a change in the type of anticancer agent can be cited.

[0148] Step (B2) may be a step of specifying a sample derived from a subject with a high possibility of having a poor prognosis when at least one selected from the group consisting of (i2) and (ii2) is satisfied.

[0149] [Test sample inspection method (3)] The third aspect of the sample inspection method is a method for evaluating whether the gynecological cancer possessed by the subject is a gynecological malignant tumor or a gynecological borderline malignant tumor. The inspection method includes a step (A3) and a step (B3). Step (A3): a step of measuring the concentration of one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid Step (B3): a step of evaluating whether the gynecological cancer is a malignant tumor or a borderline malignant tumor based on the concentration of the free fatty acid in the sample obtained in step (A3)

[0150] In the third embodiment of the sample inspection method, the sample for inspection is derived from a human female with gynecological cancer. As the gynecological cancer possessed by the subject, the cancer described in the first embodiment can be cited. For example, the subject is a human female who has been pre-diagnosed as having gynecological cancer. Alternatively, it can also be a subject determined to have gynecological cancer by the inspection method of the first embodiment described above.

[0151] The gynecological malignant tumor is preferably at least one selected from the group consisting of ovarian malignant tumor, uterine body malignant tumor and cervical malignant tumor, and more preferably ovarian malignant tumor. The gynecological borderline malignant tumor is preferably at least one selected from the group consisting of ovarian borderline malignant tumor, uterine body borderline malignant tumor and cervical borderline malignant tumor, and preferably ovarian borderline malignant tumor. The evaluation in the inspection method involved in the third aspect may be, for example, whether the ovarian cancer of the evaluation subject is an ovarian malignant tumor or an ovarian borderline malignant tumor. It may also be whether the uterine body cancer of the evaluation subject is a uterine body malignant tumor or a uterine body borderline malignant tumor. It may also be whether the cervical cancer of the evaluation subject is a cervical malignant tumor or a cervical borderline malignant tumor.

[0152] Palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid are also collectively referred to as "free fatty acids of group A3".

[0153] Step (A3) can be performed in the same manner as step (A1) in the first embodiment, except that a sample derived from a subject having gynecological cancer is used. The measurement may be performed in vitro.

[0154] In step (A3), the number of types of free fatty acids to be measured may be one or more of the free fatty acids selected from group A3, may be two or more, may be three or more, may be four or more, may be five or more, may be six or more, may be seven or more, may be eight or more, may be nine or more, may be ten or more, may be eleven or more, may be twelve or more, may be thirteen or more, may be fourteen. The free fatty acids to be measured may be arbitrarily selected from the free fatty acids in group A3.

[0155] When the gynecological cancer is ovarian cancer, in step (A3), one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid and lignoceric acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these free fatty acids. When the gynecological cancer is ovarian cancer, in step (A3), the concentrations of the ten free fatty acids are preferably measured.

[0156] When gynecological cancer is uterine corpus cancer or cervical cancer, in step (A3), one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenal acid and docosapentaenoic acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, or twelve free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these free fatty acids. When gynecological cancer is uterine corpus cancer or cervical cancer, in step (A3), the concentrations of the twelve free fatty acids are preferably measured.

[0157] When gynecological cancer is uterine cancer, in step (A3), one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, or eight free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these free fatty acids. When gynecological cancer is uterine cancer, in step (A3), the concentrations of the eight free fatty acids are preferably measured.

[0158] When the gynecological cancer is cervical cancer, in step (A3), one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid can be used as the measurement object. Two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, or twelve free fatty acids in the free fatty acids can be measured. The free fatty acids of the measurement object can be arbitrarily selected from these free fatty acids. When the detection object is cervical cancer, in step (A3), the concentrations of the twelve free fatty acids are preferably measured.

[0159] In step (B3), when at least one selected from the group consisting of the following (i3) and (ii3) is satisfied, it is evaluated that the gynecological cancer possessed by the subject is highly likely to be a malignant tumor. (i3) the concentration of at least one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid and palmitic acid in the sample is higher than a predetermined threshold value; (ii3) The concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid in the sample is lower than a predetermined threshold value.

[0160] The "predetermined threshold value" in (i3) and (ii3) above is a value set for each free fatty acid. The predetermined threshold value of the concentration of each free fatty acid can be a critical value obtained by obtaining the concentration of each free fatty acid based on multiple samples from patients with gynecological malignancies and multiple samples from patients with gynecological borderline malignancies, and performing statistical processing on the concentrations of these free fatty acids.

[0161] In the case where gynecological cancer is ovarian cancer, as the free fatty acid used in the evaluation, it is preferably selected from more than one of the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenal acid, nervonic acid and lignoceric acid. The free fatty acid used to calculate the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9 or 10 of the free fatty acids. The free fatty acid used to evaluate can be arbitrarily selected from these free fatty acids. The free fatty acid used to evaluate is preferably the 10 kinds of free fatty acids.

[0162] In the case where gynecological cancer is uterine corpus cancer or cervical cancer, as the free fatty acid for evaluation, preferably be selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenal acid and docosapentaenoic acid. The free fatty acid for evaluation can be more than 3 kinds, more than 4 kinds, more than 5 kinds, more than 6 kinds, more than 7 kinds, more than 8 kinds, more than 9 kinds, more than 10 kinds, more than 11 kinds or 12 kinds of the free fatty acid. The free fatty acid for evaluation can be selected arbitrarily from these free fatty acids. In the case where gynecological cancer is uterine corpus cancer or cervical cancer, the free fatty acid for evaluation is preferably the 12 kinds of free fatty acids.

[0163] In the case where gynecological cancer is uterine cancer, as the free fatty acid for evaluation, it is preferably selected from more than one of the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid. The free fatty acid for calculating the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7 or 8 of the free fatty acids. The free fatty acid for evaluation can be arbitrarily selected from these free fatty acids. The free fatty acid for evaluation is preferably the 8 kinds of free fatty acids.

[0164] In the case where gynecological cancer is cervical cancer, as the free fatty acid for evaluation, it is preferably selected from more than one of the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid. The free fatty acid for calculating the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9, more than 10, more than 11 or 12 of the free fatty acids. The free fatty acid for evaluation can be arbitrarily selected from these free fatty acids. The free fatty acid for evaluation is preferably the 12 kinds of free fatty acids.

[0165] Step (B3) may also be a step of calculating an FFA score, and when the FFA score is higher than a predetermined threshold value, evaluating that the gynecological cancer possessed by the subject is likely to be a malignant tumor. The FFA score is calculated by the same method as described in the first embodiment. That is, the FFA score is calculated as the total value of the scores of the free fatty acids set in the same manner as in (i-1) and (ii-1) above.

[0166] In step (B3), the predetermined threshold value set in the same manner as in (i-1) and (ii-1) above may be obtained as described above, or the same threshold value as in the inspection method of the first embodiment may be used. The predetermined threshold value for the FFA score may be a critical value obtained by calculating the FFA scores for a plurality of samples derived from patients with gynecological malignancies and a plurality of samples derived from patients with gynecological borderline malignancies and performing statistical processing on the FFA scores.

[0167] The predetermined threshold value about the FFA score also depends on the number of species of free fatty acids used in the calculation of the FFA score. For example, when the number of species of free fatty acids is set to n, more than 0.2n and less than 0.6n, more than 0.3n and less than 0.6n, or more than 0.4n and less than 0.6n, etc. can be cited. As the number of species of free fatty acids used in the calculation of the FFA score, for example, more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9 or more than 10 species of the free fatty acids in the above-mentioned A3 group can be cited. The free fatty acids used in the calculation of the FFA score can be arbitrarily selected from the free fatty acids of the A3 group.

[0168] In the case where gynecological cancer is ovarian cancer, as the free fatty acid for calculating the FFA score, it is preferably selected from more than one of the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenal acid, nervonic acid and lignoceric acid. The free fatty acid for calculating the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9 or 10 of the free fatty acids. The free fatty acid for calculating the FFA score can be arbitrarily selected from these free fatty acids. In the case where gynecological cancer is ovarian cancer, in the calculation of the FFA score, it is preferred to use the 10 free fatty acids. When using the 10 free fatty acids, as the predetermined threshold value of the FFA score, for example, more than 4 and less than 6 can be cited.

[0169] In the case where gynecological cancer is uterine corpus cancer or cervical cancer, as the free fatty acid for calculating the FFA score, it is preferably selected from more than one of the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenal acid and docosapentaenoic acid. The free fatty acid for calculating the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9, more than 10, more than 11 or 12 kinds of the free fatty acid. The free fatty acid for calculating the FFA score can be arbitrarily selected from these free fatty acids. In the case where gynecological cancer is uterine corpus cancer or cervical cancer, in the calculation of the FFA score, it is preferred to use the 12 kinds of free fatty acids. When using the 12 kinds of free fatty acids, as the predetermined threshold value of the FFA score, the same threshold value as above can be cited.

[0170] In the case where gynecological cancer is uterine cancer, as the free fatty acid for calculating the FFA score, it is preferably selected from more than one of the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid. The free fatty acid for calculating the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7 or 8 of the free fatty acids. The free fatty acid for calculating the FFA score can be arbitrarily selected from these free fatty acids. In the case where gynecological cancer is uterine cancer, the 8 kinds of free fatty acids are preferably used in the calculation of the FFA score. When using the 8 kinds of free fatty acids, as the predetermined threshold value of the FFA score, the same threshold value as above can be listed.

[0171] In the case where gynecological cancer is cervical cancer, as the free fatty acid for calculating the FFA score, it is preferably selected from more than one of the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid. The free fatty acid for calculating the FFA score can be more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9, more than 10, more than 11 or 12 kinds of the free fatty acid. The free fatty acid for calculating the FFA score can be arbitrarily selected from these free fatty acids. In the case where gynecological cancer is cervical cancer, in the calculation of the FFA score, it is preferred to use the 12 kinds of free fatty acids. When using the 12 kinds of free fatty acids, as the predetermined threshold value of the FFA score, the same threshold value as above can be cited.

[0172] In the inspection method according to the third aspect, when the FFA score is used, the following examples can be given as specific examples of the step (A3) and the step (B3). In step (A3), the concentrations of 10 free fatty acids consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, and lignoceric acid in a sample derived from a subject are measured. In step (B3), when the FFA score of 10 free fatty acids consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid and lignoceric acid is higher than a predetermined threshold, it is judged that the gynecological cancer suffered by the subject is likely to be a malignant tumor. In this case, the threshold value of the total score is preferably 4 or more and 6 or less, and more preferably 4.5 or more and 5 or less.

[0173] According to the sample inspection method of the third aspect described above, it is possible to evaluate whether the gynecological cancer possessed by the subject is a gynecological malignant tumor or a gynecological borderline malignant tumor. Borderline malignant tumors are tumors with intermediate properties between malignant tumors and benign tumors. Borderline malignant tumors of the ovary do not metastasize through the blood circulation or lymphatic circulation like malignant tumors, so the prognosis is not as poor as that of malignant tumors. That is, borderline malignant tumors of the ovary have clinical characteristics such as no metastasis although the lesions occasionally extend into the abdominal cavity. In addition, the growth rate of tumors is usually different between borderline malignant tumors and malignant tumors. Malignant tumors grow rapidly, sow seeds, and metastasize, so surgical treatment is required as quickly as possible. On the other hand, the tumor growth of borderline malignant tumors is slow, and there is no need for emergency surgery. Based on the above-mentioned differences in characteristics, non-invasively distinguishing whether a tumor is a borderline malignant tumor or a malignant tumor through a blood test before treatment (at the time of diagnosis) is extremely useful for deciding treatment guidelines (waiting time until surgery, surgical method, degree of surgical invasiveness, etc.). Currently, the tumor is identified by pathological diagnosis during surgery. However, the third-type sample inspection method can non-invasively evaluate whether the gynecological cancer a subject has is a borderline malignant tumor or a malignant tumor before surgery.

[0174] In one embodiment, the present invention provides a method for treating gynecological cancer, comprising: (a3) ​​determining a subject with a high possibility of having a gynecological borderline malignancy or a subject with a high possibility of having a gynecological borderline malignancy through a third-party sample inspection method; and (b3) treating the gynecological borderline malignancy in the subject with a high possibility of having a gynecological borderline malignancy, and treating the gynecological malignancy in the subject with a high possibility of having a gynecological borderline malignancy. In the step (a3), the sample testing method of the third embodiment is carried out as described above to identify a subject with a high possibility of having a gynecological borderline malignant tumor or a subject with a high possibility of having a gynecological malignant tumor. In the step (b3), in the subject determined in the step (a3), if the subject has a high possibility of having a gynecological borderline malignant tumor, treatment of the gynecological borderline malignant tumor is performed. As treatment of gynecological borderline malignant tumor, surgical treatment can be mentioned, preferably removal of abdominal organs (i.e., uterus, ovary, fallopian tube and greater omentum), more preferably removal of only abdominal organs. In addition, when the subject is highly likely to have a gynecological malignancy, treatment of the gynecological malignancy is performed. As treatment of gynecological malignancy, surgical treatment can be mentioned, and abdominal visceral removal and lymph node dissection are preferred.

[0175] Step (B3) may be a step of specifying a sample derived from a subject having a high possibility that the gynecological cancer is a malignant tumor when at least one selected from the group consisting of (i3) and (ii3) is satisfied.

[0176] [Reagent test kit] The fourth embodiment of the gynecological cancer and precancerous lesions inspection kit comprises a reagent for measuring one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, docosapentaenoic acid and arachidonic acid. The kit according to the present embodiment is used for at least one purpose selected from the group consisting of the following (a) to (d). (a) evaluating the possibility that the subject has one or more types of cancer selected from the group consisting of gynecological cancer and precancerous lesions thereof; (b) evaluating the possibility that a subject with gynecological cancer will have a poor prognosis; (c) whether the gynecological cancer the subject has is a malignant tumor or a borderline malignant tumor; (d) evaluating whether a subject having a high probability of having one or more selected from the group consisting of gynecological cancer and gynecological cancer precancerous lesions has gynecological cancer or gynecological cancer precancerous lesions.

[0177] The subject in (a) above is a human female. As gynecological cancers that the subject may have, the cancers described in the first embodiment can be cited. As gynecological cancers that the subject may have, preferably gynecological malignant tumors. As gynecological malignant tumors, preferably at least one selected from the group consisting of ovarian malignant tumors, uterine body malignant tumors and cervical malignant tumors, more preferably ovarian malignant tumors. As gynecological borderline malignant tumors, preferably at least one selected from the group consisting of ovarian borderline malignant tumors, uterine body borderline malignant tumors and cervical borderline malignant tumors, more preferably ovarian borderline malignant tumors. Examples of gynecological cancer precancerous lesions that a subject may have include the lesions described in the first aspect.

[0178] The subject in (b) above is a human female with gynecological cancer. As the gynecological cancer, the cancer described in the first embodiment can be cited. As the gynecological cancer, it is preferably a gynecological malignant tumor, more preferably one or more selected from the group consisting of ovarian malignant tumors, uterine body malignant tumors and cervical malignant tumors, and further preferably ovarian malignant tumors.

[0179] The subject in (c) above is a human female with gynecological cancer. As the gynecological cancer, the cancer described in the first embodiment can be cited. As the gynecological malignant tumor, it is preferably at least one selected from the group consisting of ovarian malignant tumors, uterine body malignant tumors and cervical malignant tumors, and more preferably ovarian malignant tumors. As the gynecological borderline malignant tumor, it is preferably at least one selected from the group consisting of ovarian borderline malignant tumors, uterine body borderline malignant tumors and cervical borderline malignant tumors, and preferably ovarian borderline malignant tumors. The reagent kit according to this aspect is suitable for use in the testing methods according to the first to third aspects described above.

[0180] The reagent for measuring free fatty acids is a reagent for measuring the concentration of free fatty acids in a sample derived from a subject. The reagent may be a reagent that specifically binds to any one of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, docosapentaenoic acid and arachidonic acid (free fatty acids of group A1). For example, the reagent for measuring free fatty acids may be an antibody that specifically binds to a specific free fatty acid, or a nucleic acid aptamer, etc.

[0181] When the free fatty acids in the sample are measured using analytical instruments such as gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, and gas chromatography, any of the above-mentioned standard samples of free fatty acids can be cited as reagents for measuring the concentration of the free fatty acids. The standard samples are used to prepare a standard curve in the determination of the free fatty acids using the analytical instruments as described above. The concentration of the free fatty acids in the sample can be calculated based on the standard curve prepared by the standard samples. The standard sample of free fatty acids may be a purified product of the free fatty acids described above. The purity of the specific free fatty acids in the standard sample of the specific free fatty acids is, for example, 96% by mass or more, 97% by mass or more, 98% by mass or more, or 99% by mass or more.

[0182] The test kit of the present embodiment only needs to contain any one or more reagents for measuring the free fatty acids of the above-mentioned A1 group, and may also contain any two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more reagents for measuring the free fatty acids of the above-mentioned A1 group. The fatty acids to be measured by the above-mentioned reagents that may be contained in the test kit of the present embodiment may be arbitrarily selected from the free fatty acids of the A1 group. Alternatively, the test kit of the present embodiment may also contain reagents for measuring all of the 15 free fatty acids of the above-mentioned A1 group.

[0183] In the case where the kit of the present embodiment is a test kit for gynecological cancer, it is preferably included to measure one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid. The kit of the present embodiment may contain 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 14 or more, or 14 reagents for measuring the free fatty acids. The fatty acid of the test object of the reagent that can be contained in the test kit of the present embodiment can be arbitrarily selected from these free fatty acids.

[0184] When the kit of the present embodiment is a test kit for ovarian cancer, it is preferred to include a reagent for measuring one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, and lignoceric acid. The kit of the present embodiment may include reagents for measuring three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten of the free fatty acids. The fatty acid to be measured by the reagent that the test kit of the present embodiment may contain may be arbitrarily selected from these free fatty acids.

[0185] The kit of the present embodiment preferably includes a reagent for measuring one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, and tetracosanoic acid when early gynecological cancer (e.g., stage I, stage II) is also used as a detection object. The kit of the present embodiment may include reagents for measuring three or more, four or more, five or more, six or more, seven or more, or eight of the free fatty acids. The fatty acid as the measurement object of the reagent that the test kit of the present embodiment may contain may be arbitrarily selected from these free fatty acids.

[0186] In the case where the kit of the present embodiment is a test kit for uterine body cancer or cervical cancer, it is preferably included for measuring one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid. The kit of the present embodiment may include reagents for measuring 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 of the free fatty acids. The fatty acid of the test object of the reagent that the test kit of the present embodiment may contain may be arbitrarily selected from these free fatty acids.

[0187] In the case where the kit of the present embodiment is a test kit for uterine cancer, it is preferably comprised of a reagent for measuring one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid. The kit of the present embodiment may comprise reagents for measuring three or more, four or more, five or more, six or more, seven or more, or eight of the free fatty acids. The fatty acid as the test object of the reagent that the test kit of the present embodiment may contain may be arbitrarily selected from these free fatty acids.

[0188] In the case where the kit of the present embodiment is a test kit for cervical cancer, it is preferably comprised of a reagent for measuring one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid. The kit of the present embodiment may comprise reagents for measuring three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, or twelve of the free fatty acids. The fatty acid to be measured by the reagent that may be contained in the test kit of the present embodiment may be arbitrarily selected from these free fatty acids.

[0189] In the case where the kit of the present embodiment is a test kit for the evaluation of (b) above, it is preferably included to measure one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid. The kit of the present embodiment may include 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, or 14 reagents for measuring the free fatty acids. The fatty acid as the measurement object of the reagent that the test kit of the present embodiment may contain may be arbitrarily selected from these free fatty acids.

[0190] In the case where the kit of the present embodiment is a test kit for evaluating the possibility of a subject with ovarian cancer becoming a poor prognosis, it is preferably included a reagent for measuring one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, and lignoceric acid. The kit of the present embodiment may include three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten reagents for measuring the free fatty acids. The fatty acid to be measured by the reagent that can be contained in the test kit of the present embodiment can be arbitrarily selected from these free fatty acids.

[0191] In the case where the kit of the present embodiment is an evaluation test kit for the possibility of a subject with uterine body cancer or cervical cancer becoming a poor prognosis, it is preferably included to measure a reagent for one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid. The kit of the present embodiment may include reagents for measuring 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 of the free fatty acids. The fatty acid of the test object of the reagent that can be contained in the test kit of the present embodiment can be arbitrarily selected from these free fatty acids.

[0192] In the case where the kit of the present embodiment is a test kit for evaluating the possibility that a subject with uterine body cancer has a poor prognosis, it is preferably provided with a reagent for measuring one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid. The kit of the present embodiment may include reagents for measuring three or more, four or more, five or more, six or more, seven or more, or eight of the free fatty acids. The fatty acid of the test object of the reagent that the test kit of the present embodiment may contain may be arbitrarily selected from these free fatty acids.

[0193] In the case where the kit of the present embodiment is a test kit for evaluating the possibility that a subject with cervical cancer has a poor prognosis, it is preferably provided with a reagent for measuring one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid. The kit of the present embodiment may include reagents for measuring three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, or twelve of the free fatty acids. The fatty acid to be measured by the reagent that may be contained in the test kit of the present embodiment may be arbitrarily selected from these free fatty acids.

[0194] In the case where the kit of the present embodiment is a test kit for the evaluation of (c) above, it is preferably included to measure one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid. The kit of the present embodiment may include 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, or 14 reagents for measuring the free fatty acids. The fatty acid to be measured by the reagent that the test kit of the present embodiment may contain may be arbitrarily selected from these free fatty acids.

[0195] In the case where the kit of the present embodiment is an evaluation test kit for evaluating the possibility of whether the ovarian cancer possessed by the evaluation subject is a malignant tumor or a borderline malignant tumor, it is preferably provided with a reagent for measuring one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, and lignoceric acid. The kit of the present embodiment may include reagents for measuring three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten of the free fatty acids. The fatty acid to be measured by the reagent that may be contained in the test kit of the present embodiment may be arbitrarily selected from these free fatty acids.

[0196] In the case where the kit of the present embodiment is an evaluation test kit for evaluating the possibility of whether the uterine body cancer or cervical cancer possessed by the evaluation object is a malignant tumor or a borderline malignant tumor, it is preferably included for measuring one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid. The kit of the present embodiment may include reagents for measuring 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 of the free fatty acids. The fatty acid of the test object of the reagent that can be contained in the test kit of the present embodiment can be arbitrarily selected from these free fatty acids.

[0197] In the case where the kit of the present embodiment is an evaluation test kit for evaluating the possibility of whether the uterine body cancer possessed by the evaluation object is a malignant tumor or a borderline malignant tumor, it is preferably included a reagent for measuring one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid and arachidic acid. The kit of the present embodiment may include reagents for measuring 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, or 8 of the free fatty acids. The fatty acid of the test object of the reagent that the test kit of the present embodiment may contain may be arbitrarily selected from these free fatty acids.

[0198] In the case where the kit of the present embodiment is an evaluation test kit for evaluating the possibility of whether the cervical cancer possessed by the evaluation object is a malignant tumor or a borderline malignant tumor, it is preferably included a reagent for measuring one or more free fatty acids selected from the group consisting of palmitic acid, palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid. The kit of the present embodiment may include reagents for measuring 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 of the free fatty acids. The fatty acid as the measurement object of the reagent that can be contained in the test kit of the present embodiment can be arbitrarily selected from these free fatty acids.

[0199] When the kit of the present embodiment is a test kit for the evaluation of (d) above, it preferably contains a reagent for measuring one or more free fatty acids selected from the group consisting of dihomo-γ-linolenic acid, docosahexaenoic acid, adrenal acid, and docosapentaenoic acid. The kit of the present embodiment may contain two or more, three or more, or four reagents for measuring the free fatty acids. The fatty acid to be measured by the reagent that the test kit of the present embodiment may contain may be arbitrarily selected from these free fatty acids.

[0200] When using analytical instruments such as gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, and gas chromatography to measure free fatty acids in a sample, the kit of this embodiment may include an internal standard substance. The internal standard substance is added to a sample derived from an object for use. According to the amount of the internal standard substance added to the sample derived from the object and the ratio of the measured value of the free fatty acids in the sample to the measured value of the internal standard substance, the concentration of the free fatty acids in the sample can be calculated. The internal standard substance can use a substance not contained in the sample derived from the object. For example, free fatty acids known not to exist in the human body can be used as the internal standard substance. The internal standard substance preferably has an extraction rate of the same degree as the free fatty acids of the measurement object relative to the solvent used when extracting free fatty acids from the sample derived from the object. As such an internal standard substance, for example, heptadecanoic acid etc. can be cited.

[0201] By using the kit of this embodiment, the concentration of the free fatty acid in the sample derived from the subject can be easily measured. Thus, the evaluation results of the first to third aspects of the testing method can be easily obtained.

[0202] [Medical products] The pharmaceutical product according to the present embodiment is a pharmaceutical product for treating or preventing gynecological cancer and is administered to a subject evaluated as having gynecological cancer by the examination method according to the first embodiment.

[0203] Examples of the gynecological cancer include the cancers described in the first aspect. The gynecological cancer is preferably a gynecological malignant tumor, more preferably a malignant tumor selected from the group consisting of an ovarian malignant tumor, a uterine body malignant tumor, and a cervical malignant tumor, and even more preferably an ovarian malignant tumor.

[0204] The pharmaceuticals involved in this embodiment are not particularly limited as long as they are effective for gynecological cancer. As pharmaceuticals for gynecological cancer, for example, known anticancer agents for the treatment of gynecological cancer can be cited. As such anticancer agents, for example, carboplatin, cisplatin, paclitaxel, docetaxel, irinotecan, adriamycin, gemcitabine, etoposide, travinone, albumin paclitaxel, tamoxifen, ifosfamide, 5-fluorouracil, doxorubicin, etc. can be cited. Alternatively, the pharmaceuticals of this embodiment can be antibody pharmaceuticals, for example, known antibody pharmaceuticals can be cited. As antibodies that can be contained in antibody pharmaceuticals, anti-VEGF antibodies such as bevacizumab can be cited.

[0205] The pharmaceutical product according to the present embodiment may contain various substances generally used for formulation, such as excipients, stabilizers, other pharmaceutically acceptable ingredients, or other pharmaceutically active ingredients.

[0206] The pharmaceutical product involved in this embodiment can be administered orally or parenterally. The pharmaceutical product of this embodiment can take various preparation forms, such as solid preparations, liquid preparations, etc., according to the administration route. For example, it can be made into an oral preparation such as a solid preparation or a liquid preparation for oral administration, or a parenteral preparation such as an injection or a drip. As a carrier that can be used for a parenteral preparation, for example, an aqueous carrier such as physiological saline, an isotonic solution containing glucose or D-sorbitol, etc. can be listed.

[0207] The dosage of the drug involved in this scheme varies according to the age, weight, disease, symptoms, etc. of the subject, and for example, the dosage per time may be about 0.1 μg-1 mg / kg per unit body weight. The dosage interval is not particularly limited, and may be, for example, once a day to several months.

[0208] [Other methods] In one embodiment, the present disclosure provides a biomarker for evaluating the presence of one or more biomarkers selected from the group consisting of gynecological cancer and precancerous lesions thereof, wherein the biomarker is selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, docosapentaenoic acid and arachidonic acid. In one embodiment, the present disclosure provides a biomarker for predicting the prognosis of a gynecological cancer patient, wherein the biomarker is selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid. In one embodiment, the present disclosure provides a biomarker for classifying gynecological malignancies and gynecological borderline malignancies, wherein the biomarker is selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid. In one embodiment, the present disclosure provides a biomarker for classifying gynecological cancer and its precancerous lesions, wherein the biomarker is selected from the group consisting of dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid.

[0209] In one embodiment, the present disclosure provides a use of a reagent in preparing a diagnostic drug for gynecological cancer and its precancerous lesions. The diagnostic drug is used for at least one diagnosis selected from the group consisting of the following (a) to (d). (a) the subject has a diagnosis of one or more possibilities selected from the group consisting of gynecological cancers and precancerous lesions thereof; (b) diagnosing the possibility that the subject having gynecological cancer will have a poor prognosis; (c) diagnosis of whether the gynecological cancer the subject has is a malignant tumor or a borderline malignant tumor; (d) a diagnosis of which of gynecological cancer and gynecological cancer precancerous lesions the subject has a high probability of having one or more selected from the group consisting of gynecological cancer and gynecological cancer precancerous lesions. The reagent is a purified product for measuring the concentration of free fatty acids in a sample derived from a subject. The free fatty acid is one or more selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, docosapentaenoic acid and arachidonic acid. The purified product may be one or more selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, docosapentaenoic acid and arachidonic acid. The purified product can be 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, or 15 of these fatty acids. The purified product can be arbitrarily selected from these fatty acids.

[0210] The preferred embodiments of the present invention have been described above, but the present invention is not limited to these embodiments. Additions, omissions, substitutions and other changes to the structure can be made without departing from the gist of the present invention. The present invention is not limited to the above description, but is only limited to the scope of the attached claims. [Example]

[0211] The present invention is described below by way of examples, but the present invention is not limited to the following examples. In the examples, ovarian cancer, uterine body cancer, and cervical cancer refer to ovarian malignant tumors, uterine body malignant tumors, and cervical malignant tumors, respectively.

[0212] [Experimental Example 1] (Expression analysis of fatty acid metabolic enzymes) The expression levels of fatty acid metabolic enzymes in normal ovarian tissues and ovarian cancer tissues were analyzed by RT-PCR.

[0213] 9 normal ovarian tissues were collected during surgery for gynecological malignancies for therapeutic purposes (prevention of metastasis, etc.), and 32 ovarian cancer tissues were collected during surgery for ovarian malignancies. Each normal ovarian tissue and ovarian cancer tissue was pulverized with a crusher, and total RNA was extracted using RNeasy Kit (QIAGEN).

[0214] cDNA synthesis was performed using ReverTra Ace qPCR RT Master Mix with gDNA Remover (manufactured by TOYOBO). Specifically, first, 1 μg of total RNA and RNase-free water were mixed into 12 μL, and the mixture was incubated at 65°C for 5 minutes to modify the RNA. Next, 4 μL of 4×DN Master Mix and gDNA Remover were added on ice, and the mixture was reacted at 37°C for 5 minutes to remove the genomic DNA. Next, 4 μL of 5×RT Master Mix II was further mixed, and a reverse transcription reaction was performed (37°C for 15 minutes, 50°C for 5 minutes, and 98°C for 5 minutes) to synthesize cDNA.

[0215] Real-time PCR was performed using the cDNA prepared above as a template using THUNDERBIRD Probe qPCR Mix (manufactured by TOYOBO Corporation). Real-time PCR was performed according to 2 -ΔCt That is, the difference between the Ct value of the target gene and glyceraldehyde3-phosphate dehydrogenase (GAPDH) was taken as the ΔCt value, and the relative quantitative value was calculated by 2 -ΔCt Each determination was performed in triplicate. The target genes are human acetyl-CoA carboxylase1(ACACA), fatty acid synthase(FASN), stearoyl-CoA desaturase1(SCD1), fatty acid desaturase1(FADS1), FADS2, elongation of very long chain fatty acid1(ELOVL1), ELOVL2, ELOVL3, ELOVL4, ELOVL5 and ELOVL6.

[0216] The results are shown in Figure 1A to Figure 1D . Figure 1A This is a graph showing the expression levels of free fatty acid metabolic enzymes in normal ovarian tissue and ovarian cancer tissue using a heat map. The heat map was created using MetaboAnalyst 5.0 software.

[0217] Figure 1B Figures 1 to 4 are graphs showing the expression levels of each fatty acid metabolism enzyme in normal ovarian tissue and ovarian cancer tissue. The comparison between the two groups of normal ovarian tissue and ovarian cancer tissue was performed by t-test, and p<0.05 was set as a significant difference. "*", "**", "***", and "****" represent p<0.05, p<0.01, p<0.001, and p<0.0001, respectively.

[0218] according to Figure 1B The results shown in Figure 2 confirmed that the expression levels of FASN and SCD1 in ovarian cancer tissues were higher than those in normal ovarian tissues. Figure 1C The results shown confirmed that the expression levels of ELOVL1, ELOVL2, ELOVL3, ELOVL4, ELOVL5, ELOVL6 and FADS1 in ovarian cancer tissues were lower than those in normal ovarian tissues. Figure 1D The results shown confirmed that ACACA and FADS2 were expressed at similar levels in normal ovarian tissue and ovarian cancer tissue. Based on the above results, it was confirmed that the expression levels of fatty acid metabolism enzymes were different in normal ovarian tissue and ovarian cancer tissue.

[0219] [Experimental Example 2] (Free fatty acid concentration in serum of patients with ovarian cancer) The amount of free fatty acids in the serum of female healthy donors and ovarian cancer patients was analyzed by gas chromatography-mass spectrometry (GC-MS).

[0220] Serum samples were obtained by collecting blood from 27 healthy adult women and 40 ovarian cancer patients (29 patients with stage I and stage II, and 11 patients with stage III and stage IV).

[0221] First, a standard curve was prepared using standard samples for measuring free fatty acids in serum. As standard samples, saturated and monosaturated fatty acid LC-MS mixture (manufactured by Cayman Chemical, #17942), polyunsaturated fatty acid LC-MS mixture (manufactured by Cayman Chemical, #17941), hexadecenoic acid (manufactured by Cayman Chemical, #9001845), vaccenic acid (manufactured by Cayman Chemical, #20023), and heptadecanoic acid (manufactured by Cayman Chemical, Nacalai Tesque) as an internal standard substance were used.

[0222] First, 20 μL of healthy donor serum and ovarian cancer patient serum were diluted with 300 μL of phosphate buffered saline (PBS) containing 100 ng of heptadecanoic acid as an internal standard. Then, the entire sample was applied to a diatomaceous earth column (trade name "SLE+400", Biotage) and eluted with 1.8 mL of dichloromethane. Next, the eluate was dried under a nitrogen stream, and the obtained dry matter was dissolved in 5 μL of pyridine, to which 30 μL of BSTFA:TMCS (99:1, ThermoFisher TS-38831) was added and mixed to trimethylsilylate all the OH groups.

[0223] Next, 2 μL of the derivatized sample was injected into GC-MS for analysis and resolution. The quantitative value was determined by comparing the peak area ratio of the internal standard substance and each free fatty acid with a pre-prepared calibration curve.

[0224] The analysis conditions of GC-MS are as follows. As a capillary column, RTx-5 MS (length 30 meters, inner diameter 0.25 mm, film thickness 0.25 μm, manufactured by Restek) was used. The oven temperature was maintained at 150°C for 1 minute, heated to 250°C at 20°C / min, heated to 280°C at 5°C / min, maintained at 280°C for 3 minutes, heated to 330°C at 20°C / min, and maintained at 330°C for 1 minute. He was used as a carrier gas. The linear velocity was set to 42.0 cm / sec. The ion source temperature, interface temperature, and vaporization chamber temperature were set to 200°C, 280°C, and 250°C, respectively.

[0225] The free fatty acids to be measured are 18 types, including palmitoleic acid, palmitic acid, γ-linolenic acid, stearidonic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, stearic acid, arachidic acid, arachidonic acid, eicosapentaenoic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, docosapentaenoic acid, neuraminic acid, and lignoceric acid.

[0226] The measurement results are shown in Figure 2A to Figure 2D . Figure 2A The figure shows the average values ​​of the above-mentioned various free fatty acid amounts in the sera of healthy donors and ovarian cancer patients as a heat map. The heat map was created using MetaboAnalyst 5.0 software.

[0227] Figure 2B to Figure 2C This is a graph showing the individual values ​​of various free fatty acids in the serum of healthy donors and ovarian cancer patients. The comparison between the two groups was performed by t-test, and p < 0.05 was set as a significant difference. "*", "**", "***", and "****" respectively indicate p < 0.05, p < 0.01, p < 0.001, and p < 0.0001.

[0228] Figure 2D The results of measuring each free fatty acid are shown in a volcano plot. Figure 2D In the graph, "Up" refers to free fatty acids whose concentration in the serum of ovarian cancer patients is significantly higher than that in the serum of healthy donors. "Down" refers to free fatty acids whose concentration in the serum of ovarian cancer patients is significantly lower than that in the serum of healthy donors. "Non-sig" refers to free fatty acids for which no significant difference in concentration was confirmed between the serum of healthy donors and the serum of ovarian cancer patients. The horizontal axis represents the ratio of the average value of the concentration of each free fatty acid in the serum of ovarian cancer patients to the average value of the concentration of each free fatty acid in the serum of healthy donors. The vertical axis represents the P value when there is no difference in the concentration of each free fatty acid in the serum of healthy donors and the concentration of each free fatty acid in the serum of ovarian cancer patients.

[0229] according to Figure 2AThe results shown in Figure 1 to Figure 2 confirmed that the concentrations of palmitoleic acid, palmitic acid, linoleic acid, α-linolenic acid, and oleic acid were higher in the serum of ovarian cancer patients than in the serum of healthy donors. In contrast, the concentrations of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, and lignoceric acid were lower in the serum of ovarian cancer patients than in the serum of healthy donors.

[0230] The free fatty acids with significant differences between the serum of healthy donors and the serum of ovarian cancer patients are summarized in Table 1. In Table 1, the record of "high" in the column of concentration difference means that the concentration of the free fatty acid in the serum of ovarian cancer patients is significantly higher than that in the serum of healthy donors. In Table 1, the record of "low" in the column of concentration difference means that the concentration of the free fatty acid in the serum of ovarian cancer patients is significantly lower than that in the serum of healthy donors.

[0231] [Table 1]

[0232] [Experimental Example 3] (Correlation analysis between fatty acid metabolic enzyme expression and free fatty acid concentration) The correlation between the expression levels of fatty acid metabolic enzymes in tissues and the free fatty acid concentration in serum was analyzed.

[0233] Figure 3A-3B Yes means Figure 1B Changes in the expression of fatty acid metabolic enzymes in the tissues shown in Figure 4 Figure 2B Figure 1 to 2 are graphs showing the correlation between changes in the free fatty acid composition in serum. Figure 1B In ~D, "HIGH" refers to metabolic enzymes that are significantly more expressed in ovarian cancer tissue than in normal ovarian tissue. Figure 1B In ~D, "LOW" refers to metabolic enzymes whose expression in ovarian cancer tissue is significantly lower than that in normal ovarian tissue. Figure 1B In ~D, "not significant" means that no significant difference in expression was found between normal ovarian tissue and ovarian cancer tissue.

[0234] Ovarian cancer tissue and serum samples were collected in pairs from 20 ovarian cancer patients at the same time, and the correlation between the expression level of SCD1 in ovarian cancer tissue and the ratio of free fatty acids, which are substrates of SCD1, to free fatty acids generated by the action of SCD1 was analyzed.

[0235] SCD1 acts on palmitic acid as a substrate to generate palmitoleic acid, and on stearic acid to generate oleic acid.

[0236] Figure 3CThe graph shows the correlation between the SCD1 gene expression level in ovarian cancer tissue and the free fatty acid ratio (produced free fatty acid / substrate free fatty acid ratio (palmitoleic acid / palmitic acid ratio)) corresponding to SCD1 activity in the serum of ovarian cancer patients. Figure 3D This is a graph showing the correlation between the SCD1 gene expression level in ovarian cancer tissue and the free fatty acid ratio (produced free fatty acid / matrix free fatty acid ratio (oleic acid / stearic acid ratio)) equivalent to SCD1 activity in the serum of ovarian cancer patients. Correlation analysis was performed using GraphPad Software version 9.0, using Pearson's correlation test, and r and p values ​​were calculated. Depend on Figures 3A to 3D The results shown indicate that changes in the expression of fatty acid metabolic enzymes in tissues are linked to changes in the concentration of free fatty acids in serum.

[0237] [Experimental Example 4] (Detection of ovarian cancer patients based on ROC analysis of free fatty acid concentration) In Experimental Example 2, Receiver operating characteristic analysis (ROC analysis) was performed using 11 free fatty acids whose concentrations were found to be significantly different between the serum of healthy female donors and ovarian cancer patients as indicators.

[0238] ROC analysis was performed on healthy donors and ovarian cancer patients using the concentration of each free fatty acid. Figure 4A The results of ROC analysis using the concentrations of palmitoleic acid, palmitic acid, linoleic acid, α-linolenic acid, and oleic acid are shown. Figure 4B The results of ROC analysis using the concentrations of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, and lignoceric acid are shown. Figure 4A In ~B, AUC and CI refer to Area Under Curve and Confidence Interval, respectively.

[0239] Among these 11 types, based on the concentrations of 10 free fatty acids, namely palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, neuraminic acid and lignoceric acid, it is clear whether the subject of the examination is a healthy donor or an ovarian cancer patient.

[0240] [Experimental Example 5] (Detection of ovarian cancer patients based on ROC analysis of FFA score) For the 10 free fatty acids with good results in the ROC analysis of Experimental Example 4, threshold values ​​were set so that the positive likelihood ratio of the ROC analysis in Experimental Example 4 was maximized. The threshold values ​​of each free fatty acid were as follows: 29.06 μEQ / L for palmitoleic acid, 326.0 μEQ / L for oleic acid, 183.1 μEQ / L for linoleic acid, 28.62 μEQ / L for α-linolenic acid, 1.926 μEQ / L for vaccenic acid, 1.924 μEQ / L for arachidic acid, 0.7085 μEQ / L for adrenic acid, 1.847 μEQ / L for docosahexaenoic acid, 0.5773 μEQ / L for lignoceric acid, and 0.0891 μEQ / L for neuraminic acid.

[0241] For the subjects analyzed in Experimental Example 2, based on the threshold values ​​of the 10 free fatty acids set above, it was determined whether the serum concentration of each free fatty acid was an abnormal value. Figure 5A The results for ovarian cancer patients are shown in Figure 5B . Figures 5A-5B In the table, “1” represents palmitoleic acid, “2” represents oleic acid, “3” represents linoleic acid, “4” represents α-linolenic acid, “5” represents vaccenic acid, “6” represents arachidic acid, “7” represents adrenic acid, “8” represents docosahexaenoic acid, “9” represents lignoceric acid, and “10” represents neuraminic acid. Regarding free fatty acids “1” to “4”, when the measured concentration is higher than the threshold value, the concentration is determined to be an abnormal value and is represented by a black circle. When the measured concentration is lower than the threshold value, the concentration is determined to be a normal value and is represented by a white circle. Regarding free fatty acids “5” to “10”, when the measured concentration is lower than the threshold value, the concentration is determined to be an abnormal value and is represented by a black circle. When the measured concentration is higher than the threshold value, the concentration is determined to be a normal value and is represented by a white circle.

[0242] according to Figures 5A-5B The number of black circles for each subject was calculated as the FFA score for each subject. Using this FFA score, ROC analysis was performed on female healthy donors and ovarian cancer patients. The results of ROC analysis are shown in Figure 5C The results of ROC analysis showed that the AUC value was 0.9963. The CI value was 0.9880-1.000. The p value was P<0.0001. This result confirmed that ovarian cancer patients can be detected with high sensitivity by using this FFA score.

[0243] [Experimental Example 6] (Free fatty acid concentration in serum of patients with early ovarian cancer) The serum concentrations of the 10 free fatty acids showing good results in the ROC analysis in Experimental Example 4 were compared for 27 female healthy donors, 29 stage I or stage II ovarian cancer patients, and 11 stage III or stage IV ovarian cancer patients.

[0244] The results of the comparison among the three groups are shown in Fig. 6A and Figure 6B In the figure, "HD" refers to female healthy donors. Comparisons among the three groups were performed using one-way ANOVA and Bonferroni's multiple comparisons tests, with p < 0.05 considered significant. "*", "**", "***", and "****" indicate p < 0.05, p < 0.01, p < 0.001, and p < 0.0001, respectively.

[0245] Depend on Fig. 6A The results shown confirmed that the concentrations of four free fatty acids, namely palmitoleic acid, linoleic acid, α-linolenic acid and oleic acid, were significantly higher in the serum of patients with stage I or stage II ovarian cancer than in the serum of healthy donors. Depend on Figure 6B The results shown confirmed that the concentrations of four free fatty acids, namely, vaccenic acid, arachidic acid, docosahexaenoic acid, and lignoceric acid, were significantly lower in the serum of patients with stage I or stage II ovarian cancer than in the serum of healthy donors. It was confirmed that the serum concentrations of the above eight free fatty acids can serve as an indicator for distinguishing healthy donors from stage I or stage II ovarian cancer patients. It should be noted that no significant differences were found in the serum concentrations of α-linolenic acid, docosahexaenoic acid, and lignoceric acid between patients with stage III or IV ovarian cancer and healthy donors.

[0246] [Experimental Example 7] (Detection of early ovarian cancer patients based on ROC analysis of free fatty acid concentration) In Experimental Example 6, receiver operating characteristic analysis (ROC analysis) was performed using the serum concentrations of eight free fatty acids for which significant differences were confirmed between the serum concentrations of healthy female donors and stage I or stage II ovarian cancer patients as indicators.

[0247] ROC analysis was performed using the concentrations of each free fatty acid in healthy donors and patients with stage I or stage II ovarian cancer. Fig. 7A The results of ROC analysis using the concentrations of palmitoleic acid, linoleic acid, α-linolenic acid, and oleic acid are shown. Figure 7BThe results of ROC analysis using the concentrations of vaccenic acid, arachidic acid, docosahexaenoic acid, and lignoceric acid are shown. Fig. 7A In ~B, AUC and CI refer to Area Under Curve and Confidence Interval, respectively.

[0248] It was determined that based on the concentrations of these eight free fatty acids, it was possible to distinguish whether the subject of the examination was a healthy donor or a patient with stage I or stage II ovarian cancer.

[0249] [Experimental Example 8] (Detection of early ovarian cancer patients based on ROC analysis of FFA score) For the 8 free fatty acids in the ROC analysis in Experimental Example 7, thresholds were set so that the positive likelihood ratio of the ROC analysis in Experimental Example 7 was maximized. The thresholds for each free fatty acid were 29.06 μEQ / L for palmitoleic acid, 326.0 μEQ / L for oleic acid, 183.1 μEQ / L for linoleic acid, 28.62 μEQ / L for α-linolenic acid, 1.930 μEQ / L for vaccenic acid, 1.953 μEQ / L for arachidic acid, 1.847 μEQ / L for docosahexaenoic acid, and 0.5773 μEQ / L for tetracosenoic acid.

[0250] For the subjects analyzed in Experimental Example 6, based on the threshold values ​​of the eight free fatty acids set above, it was determined whether the serum concentration of each free fatty acid was an abnormal value. Fig. 8A shown. Figure 8B Results are shown for patients with stage I or stage II ovarian cancer. Fig. 8A In ~B, "1" represents palmitoleic acid, "2" represents oleic acid, "3" represents linoleic acid, "4" represents α-linolenic acid, "5" represents vaccenic acid, "6" represents arachidic acid, "7" represents docosahexaenoic acid, and "8" represents lignoceric acid. Regarding the free fatty acids of "1" to "4", when the measured concentration is higher than the threshold value, the concentration is determined to be an abnormal value and is represented by a black circle. When the measured concentration is lower than the threshold value, the concentration is determined to be a normal value and is represented by a white circle. Regarding the free fatty acids of "5" to "8", when the measured concentration is lower than the threshold value, the concentration is determined to be an abnormal value and is represented by a black circle. When the measured concentration is higher than the threshold value, the concentration is determined to be a normal value and is represented by a white circle.

[0251] according to Figures 8A-8B The number of black circles for each subject was calculated as the FFA score for each subject. Using this FFA score, ROC analysis was performed on healthy female individuals and stage I and stage II ovarian cancer patients. The results are shown in Figure 8CThe result of ROC analysis was that the AUC value was 0.9955. The 95% CI value was 0.9849-1.000. The p value was P<0.0001. From this result, it was confirmed that by using this FFA score, stage I or stage II ovarian cancer patients can be detected with high sensitivity.

[0252] The threshold of FFA score is set so that Figure 8C The positive likelihood ratio of the ROC analysis in becomes the largest. The threshold of the FFA score is 1.5.

[0253] Using the above-mentioned threshold value of the FFA score, the detection sensitivity based on the FFA score was compared with the detection sensitivity based on the blood concentration of CA125 for stage I or II ovarian cancer. Fig.8D The FFA scores (left graph) and the blood concentrations of CA125 (right graph) of patients with stage I or stage II ovarian cancer are shown. Fig.8D In the figure, “×” indicates clear cell ovarian cancer, and “●” indicates ovarian cancer other than clear cell cancer.

[0254] according to Fig.8D According to the analysis results, when the threshold value of the FFA score is 1.5, the detection sensitivity of patients with stage I or stage II ovarian cancer is 100%. When the threshold value of the blood concentration of CA125 is set to 35U / mL used in actual clinical practice, the detection sensitivity of patients with stage I or stage II ovarian cancer is 76%. From these results, it can be confirmed that by using the FFA score calculated from the 8 free fatty acid values, patients with stage I or stage II ovarian cancer can be detected with high sensitivity compared to the examination using the blood concentration of CA125. Furthermore, the FFA score confirms that it can also detect ovarian cancer of the clear cell type, which is a tissue type that is difficult to detect in CA125, with high sensitivity.

[0255] [Experimental Example 9] (Prognosis prediction for patients with ovarian cancer)

[0256] For the 40 ovarian cancer patients who were analyzed in Experimental Example 5, the median value of the FFA scores of these ovarian cancer patients was calculated from the FFA scores calculated in Experimental Example 5. Next, for the above 40 ovarian cancer patients, patients whose FFA scores were higher than the median value were classified into the "FFA score high" group, and patients whose FFA scores were lower than the median value were classified into the "FFA score low" group. In addition, the serum sample used for the calculation of the FFA score in Experimental Example 5 was based on a blood sample collected before the start of treatment. For patients in the FFA score high group and patients in the FFA score low group, the prognosis was tracked separately, and the change in the progression-free survival rate in each patient group was investigated. The results are shown in Fig. 9 . Fig. 9In the data, the X-axis is the number of days from the start of treatment, and the Y-axis is the progression-free survival rate. The Log-rank test was used to test for significant differences.

[0257] It was confirmed that the FFA score can be used as an indicator to predict the treatment prognosis of ovarian cancer patients.

[0258] [Experimental Example 10] (Detection of borderline ovarian malignant tumors based on ROC analysis of FFA score) Blood was collected from 7 patients with ovarian borderline malignant tumors to obtain serum samples. Then, the serum concentration of each free fatty acid was measured by GC-MS using the same method as in Experimental Example 2. The free fatty acids to be measured were the 10 free fatty acids analyzed in Experimental Example 5. Then, the FFA score of each patient with ovarian borderline malignant tumor was calculated using the same method as in Experimental Example 5.

[0259] The threshold values ​​of each free fatty acid used in the calculation of the FFA score were the same as those set in Experimental Example 5. That is, palmitoleic acid was 29.06 μEQ / L, oleic acid was 326.0 μEQ / L, linoleic acid was 183.1 μEQ / L, α-linolenic acid was 28.62 μEQ / L, vaccenic acid was 1.926 μEQ / L, arachidic acid was 1.924 μEQ / L, adrenic acid was 0.7085 μEQ / L, docosahexaenoic acid was 1.847 μEQ / L, lignoceric acid was 0.5773 μEQ / L, and neuraminic acid was 0.0891 μEQ / L.

[0260] Fig. 10A The graph shows the results of determining whether the serum concentration of each free fatty acid is an abnormal value based on the above threshold value for patients with borderline malignant tumors.

[0261] Use according to Fig. 10A The FFA score of patients with borderline ovarian malignancies was calculated based on the results of Figure 5B The FFA scores of ovarian cancer patients (ovarian malignant tumor patients) calculated by the results were analyzed by ROC analysis. Fig. 10B As a result of ROC analysis, the AUC value was 0.8946. The 95% CI value was 0.8003-1.000. The p value was P<0.0001. From these results, it was confirmed that the use of FFA score can distinguish between patients with ovarian borderline malignant tumors and patients with ovarian cancer (patients with ovarian malignant tumors) with high sensitivity.

[0262] [Experimental Example 11] (Analysis on uterine cancer and cervical cancer) Blood was collected from 27 healthy adult women, 66 uterine cancer patients, and 45 cervical cancer patients to obtain serum samples. Then, the serum concentration of each free fatty acid was measured by GC-MS in the same manner as in Experimental Example 2.

[0263] The measurement results are shown in Fig.11A ~ B. Comparisons among the three groups were performed using one-way ANOVA and Bonferroni's multiple comparisons test, with p < 0.05 considered significant. "**" and "****" indicate p < 0.01 and p < 0.0001, respectively.

[0264] It was confirmed that the concentrations of palmitoleic acid, palmitic acid, linoleic acid, α-linolenic acid, and oleic acid were higher in the serum of patients with uterine corpus cancer and cervical cancer than in the serum of healthy donors. It was confirmed that the concentrations of vaccenic acid, arachidic acid, and stearic acid were lower in the serum of patients with uterine corpus cancer and cervical cancer than in the serum of healthy donors. Based on these results, it can be seen that the serum concentrations of these free fatty acids can be used as an indicator to distinguish whether the subject of the examination is a healthy donor or a patient with uterine corpus cancer or cervical cancer.

[0265] [Experimental Example 12] (Free fatty acid concentration in serum of patients with early cervical cancer) Blood was collected from 27 healthy adult women, 29 stage I cervical cancer patients, and 23 stage II or III cervical cancer patients to obtain serum samples. Then, the serum concentration of each free fatty acid was measured by GC-MS in the same manner as in Experimental Example 2.

[0266] The measurement results are shown in Fig. 12A ~ C. Comparisons among the three groups were performed using one-way ANOVA and Bonferroni's multiple comparisons test, with p < 0.05 considered significant. "*", "**", "***" and "****" indicate p < 0.05, p < 0.01, p < 0.001 and p < 0.0001, respectively.

[0267] It was confirmed that the concentrations of palmitoleic acid, linoleic acid, α-linolenic acid, and oleic acid were significantly higher in the serum of patients with stage I cervical cancer or patients with stage II or III cervical cancer than in the serum of healthy donors. On the other hand, it was confirmed that the concentrations of vaccenic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid were significantly lower in the serum of patients with stage I cervical cancer or patients with stage II or III cervical cancer than in the serum of healthy donors. It was confirmed that the serum concentrations of these free fatty acids can serve as indicators for distinguishing healthy donors from stage I, stage II, or stage III cervical cancer patients.

[0268] Furthermore, the threshold value of each free fatty acid concentration was set to the value shown in Table 2, and ROC analysis was performed in healthy donors and stage I cervical cancer patients using the concentration of each free fatty acid. The results are shown in Fig. 12A The AUC of all free fatty acids was above 0.7. It was confirmed that the serum concentration of these free fatty acids can be used as an indicator to distinguish whether the subject of the examination is a healthy donor or a stage I cervical cancer patient.

[0269] [Table 2]

[0270] [Experimental Example 13] (Analysis of free fatty acid concentration ratio in serum of patients with early cervical cancer) The concentration ratio of 10 free fatty acids was used as an indicator to verify whether it was possible to distinguish whether the examined subject was a healthy donor or a cervical cancer patient.

[0271] For the serum sample of Experimental Example 12, the ratio of docosapentaenoic acid to oleic acid (docosapentaenoic acid / oleic acid) was calculated. Fig.13A The comparison among the three groups was performed by one-way ANOVA and Bonferroni's multiple comparisons test, and p < 0.05 was considered to be a significant difference. "*", "**", "***" and "****" indicate p < 0.05, p < 0.01, p < 0.001 and p < 0.0001, respectively.

[0272] It was confirmed that the ratio (docosapentaenoic acid / oleic acid) was lower in the serum of stage I cervical cancer patients or stage II or III cervical cancer patients than in the serum of healthy donors. It was confirmed that the ratio (docosapentaenoic acid / oleic acid) can be an indicator for distinguishing healthy donors from stage I, stage II, or stage III cervical cancer patients.

[0273] Furthermore, the threshold of the ratio (docosapentaenoic acid / oleic acid) was set to the value shown in Table 2, and ROC analysis was performed using the ratio (docosapentaenoic acid / oleic acid) in healthy donors and stage I cervical cancer patients. The results are shown in Fig.13A The ROC analysis result was AUC = 0.981, P < 0.0001. That is, it was confirmed that using the ratio (docosapentaenoic acid / oleic acid) as an indicator can distinguish whether the object of examination is a healthy donor or a stage I cervical cancer patient with higher accuracy than using either docosapentaenoic acid or oleic acid as an indicator.

[0274] Next, for the 29 stage I cervical cancer patients examined in Experimental Example 12, the detection sensitivity based on the ratio (docosapentaenoic acid / oleic acid) was calculated and compared with the cancer detection sensitivity based on existing diagnostic markers. The results are shown in Fig. 13B .

[0275] When the threshold value of the ratio (docosapentaenoic acid / oleic acid) was set to 0.004433, the detection sensitivity of stage I cervical cancer patients was 93.1%. When the threshold value of the blood concentration of SCC was set to 1.5 ng / mL, which is used in actual clinical practice, the detection sensitivity of stage I cervical cancer patients was 68.2%. When the threshold value of the blood concentration of CEA was set to 5.0 ng / mL, which is used in actual clinical practice, the detection sensitivity of stage I cervical cancer patients was 4.2%.

[0276] These results confirmed that by using the ratio (docosapentaenoic acid / oleic acid), stage I cervical cancer patients can be detected with high sensitivity compared with examination using blood concentrations of SCC or CEA, which are existing diagnostic markers.

[0277] [Experimental Example 14] (Analysis of patients with cervical precancerous lesions) Blood was collected from 27 healthy adult females and 17 patients with cervical precancerous lesions (cervical abnormal formation: CIN) to obtain serum samples. Then, the serum concentration of each free fatty acid was measured by GC-MS in the same manner as in Experimental Example 2.

[0278] The measurement results are shown in Fig.14A B. Comparisons among the three groups were performed using one-way ANOVA and Bonferroni's multiple comparisons test, and p < 0.05 was considered significant. "*", "**", "***" and "****" indicate p < 0.05, p < 0.01, p < 0.001 and p < 0.0001, respectively.

[0279] The concentrations of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, and arachidonic acid were confirmed to be significantly higher in the serum of CIN patients than in the serum of healthy donors. On the other hand, the concentration of arachidic acid was confirmed to be significantly lower in the serum of CIN patients than in the serum of healthy donors. It was confirmed that the serum concentrations of these free fatty acids can serve as indicators for distinguishing healthy donors from CIN patients.

[0280] Furthermore, the threshold value of each free fatty acid concentration was set to the value shown in Table 3, and ROC analysis was performed in healthy donors and CIN patients using the concentration of each free fatty acid. The results are shown in Fig.14A~ B. The AUC of all free fatty acids was above 0.7. It was found that the serum concentration of these free fatty acids could be used as an index to distinguish whether the subject of the examination was a healthy donor or a CIN patient.

[0281] [Table 3]

[0282] [Experimental Example 15] (Analysis of CIN patients and cervical cancer patients) For the serum samples of 17 CIN patients collected in Experimental Example 14 and the serum samples of 52 cervical cancer patients collected in Experimental Example 12, it was verified whether it was possible to distinguish whether the examined object was a CIN patient or a cervical cancer patient.

[0283] The analysis results are shown in Fig.15 The comparison among the three groups was performed by one-way ANOVA and Bonferroni's multiple comparisons test, and p < 0.05 was considered to be significantly different. "***" and "****" indicate p < 0.001 and p < 0.0001, respectively.

[0284] It was confirmed that the concentrations of dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid in the sera of cervical cancer patients were significantly lower than those in the sera of CIN patients. It was confirmed that the serum concentrations of these free fatty acids can serve as an indicator for distinguishing CIN patients from cervical cancer patients.

[0285] Furthermore, the threshold values ​​of the concentrations of the free fatty acids were set to the values ​​shown in Table 4, and ROC analysis was performed on CIN patients and cervical cancer patients using the concentrations of the free fatty acids. The results are shown in Fig.15 The AUC of all free fatty acids was above 0.7. It was confirmed that the serum concentration of these free fatty acids can be used as an index to distinguish whether the subject of the examination is a CIN patient or a cervical cancer patient.

[0286] [Table 4] Possibility of industrial application

[0287] According to the present invention, a simple and highly accurate inspection method for detecting one or more selected from a group consisting of gynecological cancers and their precancerous lesions, predicting the prognosis of gynecological cancers, and classifying gynecological malignant tumors and gynecological borderline malignant tumors, a pharmaceutical product for treating a subject determined to have gynecological cancer by the inspection method, and a kit can be provided. The present invention is useful for early diagnosis of gynecological cancers or their precancerous lesions.

Claims

1. A method for inspecting a sample, wherein: The sample inspection method includes: step (A1), in which the concentration of one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, docosapentaenoic acid and arachidonic acid in a sample derived from a subject is measured; and step (B1), in which the possibility of the subject having one or more free fatty acids selected from the group consisting of gynecological cancer and precancerous lesions thereof is evaluated based on the concentration of the free fatty acids in the sample obtained in step (A1), In the step (B1), when at least one selected from the group consisting of the following (i1), (ii1) and (iii1) is satisfied, it is evaluated that the subject has a high possibility of having one or more selected from the group consisting of gynecological cancer and precancerous lesions thereof: (i1) the concentration of at least one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid and arachidonic acid in the sample is higher than a predetermined threshold value; (ii1) the concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid in the sample is below a predetermined threshold; (iii1) The concentration ratio of docosapentaenoic acid to oleic acid is lower than a predetermined threshold value.

2. A method for inspecting a sample, wherein: The sample inspection method includes: a step (A2), in which the concentration of one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid is measured in a sample derived from a subject having gynecological cancer; and a step (B2), in which the possibility of the subject having a poor prognosis is evaluated based on the concentration of the free fatty acids in the sample obtained in the step (A2), In the step (B2), when at least one selected from the group consisting of the following (i2) and (ii2) is satisfied, it is evaluated that the subject has a high possibility of having a poor prognosis: (i2) the concentration of at least one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid and palmitic acid in the sample is higher than a predetermined threshold; (ii2) the concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid in the sample is lower than a predetermined threshold value.

3. A method for inspecting a sample, wherein: The sample inspection method comprises: a step (A3), in which the concentration of one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid is measured in a sample derived from a subject having gynecological cancer; and a step (B3), in which, based on the concentration of the free fatty acids in the sample obtained in the step (A3), whether the gynecological cancer possessed by the subject is a malignant tumor or a borderline malignant tumor is evaluated, In the step (B3), when at least one selected from the group consisting of the following (i3) and (ii3) is satisfied, it is evaluated that the possibility that the gynecological cancer is a malignant tumor is high: (i3) the concentration of at least one free fatty acid selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid and palmitic acid in the sample is higher than a predetermined threshold; (ii3) the concentration of at least one free fatty acid selected from the group consisting of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid and docosapentaenoic acid in the sample is lower than a predetermined threshold value.

4. The method for inspecting a sample according to claim 1, wherein: The sample inspection method further includes a step (C1), in which, based on the concentration of the free fatty acid in the sample obtained in the step (A1), it is evaluated whether the subject determined by the step (B1) to have a high probability of having one or more selected from the group consisting of gynecological cancer and its precancerous lesions has gynecological cancer or gynecological cancer precancerous lesions, In the step (C1), when the following (Ci) is satisfied, it is evaluated that the subject is highly likely to have gynecological cancer: (Ci) the concentration of at least one free fatty acid selected from the group consisting of dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid and docosapentaenoic acid in the sample is lower than a predetermined threshold value.

5. The method for inspecting a sample according to any one of claims 1 to 4, wherein: The gynecological cancer comprises at least one selected from the group consisting of ovarian cancer, uterine cancer and cervical cancer.

6. The method for inspecting a sample according to claim 1, wherein: The gynecological cancer is a stage I or stage II gynecological malignancy.

7. The sample inspection method according to claim 6, wherein: The step (A1) is a step of measuring the concentration of one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid and stearic acid. The free fatty acid in (i1) of the step (B1) is at least one selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid and palmitic acid, and the free fatty acid in (ii1) is at least one selected from the group consisting of vaccenic acid, arachidic acid and stearic acid.

8. The method for inspecting a sample according to any one of claims 1 to 4, wherein: The sample is a blood sample.

9. A test kit for gynecological cancer and its precancerous lesions, wherein: The test kit comprises a reagent for measuring one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, docosapentaenoic acid and arachidonic acid, The test kit is used for at least one purpose selected from the group consisting of the following (a) to (d): (a) evaluating the possibility that the subject has one or more types of cancer selected from the group consisting of gynecological cancer and precancerous lesions thereof; (b) assessing the likelihood that a subject with gynecological cancer will have a poor prognosis; (c) whether the gynecological cancer the subject has is a malignant tumor or a borderline malignant tumor; (d) evaluating whether a subject having a high probability of having one or more selected from the group consisting of gynecological cancer and gynecological cancer precancerous lesions has gynecological cancer or gynecological cancer precancerous lesions.

10. The gynecological cancer examination kit according to claim 9, wherein The test kit is used in the test method for a sample according to any one of claims 1 to 4.

11. A pharmaceutical product for treating or preventing gynecological cancer, wherein: The pharmaceutical product is administered to a subject evaluated as having gynecological cancer by the examination method according to claim 1.

Images