Method for examining a sample, kit for examining gynecological cancer and its precancerous lesions, and medicine

Abstract

To provide: a simple and highly accurate sample test method which can be applied to the evaluation of the presence or absence of at least one selected from the group consisting of a gynecological cancer and a precancerous lesion thereof, the evaluation of the prognosis of a gynecological cancer patient, or the evaluation of the degree of malignancy of a gynecological cancer; a test kit which can be used for the test method; and a medicine for treating or preventing a gynecological cancer which is administered to a subject detected by the test method.SOLUTION: A sample test method is employed, the method comprising a step (A) for measuring the concentration of a free fatty acid in a sample from a subject, and a step (B) for evaluating the possibility that the subject has a gynecological cancer on the basis of the concentration of the free fatty acid obtained in the step (A).SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a method for examining a sample, a kit for examining gynecological cancer and its precancerous lesions, and a medicine.

Background Art

[0002] Since ovarian cancer is a tumor that occurs in the abdominal cavity, it is difficult to exhibit symptoms and it is extremely difficult to diagnose pathologically by biopsy. In addition, for other gynecological cancers such as endometrial cancer and cervical cancer, no serum tumor marker for early diagnosis has been established, and currently, there are still many cases where they are accidentally discovered due to abnormal bleeding or the like.

[0003] As an ovarian cancer tumor marker, for example, CA125 is known (see, for example, Patent Document 1). However, since the blood concentration of CA125 is not necessarily high in early ovarian cancer, mucinous ovarian cancer, and ovarian clear cell carcinoma, these cancers may not be positively diagnosed. In addition, since the blood concentration of CA125 may also increase in benign diseases and inflammatory diseases, the determination based on the blood concentration of CA125 has a high false positive rate. Therefore, CA125 is insufficient as a tumor marker for ovarian cancer.

[0004] In recent years, as a cancer diagnosis method, a method for detecting specific exosomes or microRNAs in peripheral blood has been proposed (see, for example, Non-Patent Document 1). However, when analyzing exosomes and microRNAs from a biological sample, operations such as ultracentrifugation and affinity separation are required. Therefore, the examination using these as tumor markers is complicated and time-consuming. In addition, there is also a problem that a certain number of false positives exist, and it has not yet been put into practical use.

Prior Art Documents

Patent Documents

[0005]

Patent Document 1

Non-Patent Documents

[0006]

Non-Patent Document 1

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, there is a need for a method that can detect gynecological cancer early with high accuracy in a simple manner.

[0008] Therefore, the present invention provides a simple and highly accurate method for examining a sample applicable to the evaluation of the presence or absence of one or more selected from the group consisting of gynecological cancers and their precancerous lesions, the evaluation of the prognosis of a gynecological cancer patient, or the evaluation of the malignancy of a gynecological cancer, a test kit usable for the examination method, and a medicament for treating or preventing a gynecological cancer to be administered to a subject detected by the examination method.

Means for Solving the Problems

[0009] The present invention includes the following aspects. [1] 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; Based on the concentration of the free fatty acid in the sample obtained in the step (A1), a step (B1) of evaluating the possibility that the subject has one or more selected from the group consisting of gynecological cancers and their precancerous lesions; In the step (B1), when at least one selected from the group consisting of the following (i1), (ii1), and (iii1) is applicable, it is evaluated that the subject has a high possibility of having one or more selected from the group consisting of gynecological cancers and their precancerous lesions. A method for examining a sample; (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 high compared to a predetermined threshold; (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 low compared to a predetermined threshold; (iii1) The concentration ratio of docosapentaenoic acid to oleic acid is low compared to a predetermined threshold.

[0010] [2] 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 from a subject having a gynecological cancer; Based on the concentration of the free fatty acid in the sample obtained in the step (A2), a step (B2) of evaluating the possibility that the subject has a poor prognosis; In the step (B2), when at least one selected from the group consisting of the following (i2) and (ii2) is applicable, it is evaluated that the subject has a high possibility of having a poor prognosis. A method for examining a sample; (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 high as compared with 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 low as compared with a predetermined threshold value.

[0011] [3] 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 a gynecological cancer; 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 acid in the sample obtained in the step (A3), and In the step (B3), a method for examining a sample, wherein when at least one of the following (i3) and (ii3) is satisfied, it is evaluated that the gynecological cancer 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 high as compared with 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 low as compared with a predetermined threshold value.

[0012] [4] For the subject determined in step (B1) to have a high likelihood of having one or more selected from the group consisting of gynecological cancers and their precancerous lesions, a step (C1) of evaluating, based on the concentration of the free fatty acids in the sample obtained in step (A1), whether the subject has any of gynecological cancers and gynecological precancerous lesions is further included. The method for examining a sample according to [1], wherein in step (C1), when the following (Ci) is satisfied, it is evaluated that the subject has a high likelihood of having a 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 low as compared with a predetermined threshold value.

[0013] [5] The method for examining a sample according to any one of [1] to [4], wherein the gynecological cancer includes at least one selected from the group consisting of ovarian cancer, endometrial cancer, and cervical cancer. [6] The method for examining a sample according to [1], wherein the gynecological cancer is a stage I or stage II gynecological malignancy. [7] 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 method for examining a sample according to [6], wherein the free fatty acid in (i1) of 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 examining a sample according to any one of [1] to [4], wherein the sample is a blood sample.

[0014] [9] 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, A test kit for gynecological cancer and its precancerous lesions, used for at least one purpose selected from the group consisting of the following (a) to (d); (a) Evaluation of the possibility that a subject has one or more selected from the group consisting of gynecological cancer and its precancerous lesions; (b) Evaluation of the possibility that a subject with gynecological cancer has a poor prognosis; (c) Evaluation of whether the gynecological cancer a subject has is a malignant tumor or a borderline malignant tumor; (d) Evaluation of which one of gynecological cancer and gynecological precancerous lesions a subject with a high possibility of having one or more selected from the group consisting of gynecological cancer and gynecological precancerous lesions has.

[10] The test kit for gynecological cancer according to [9], which is used in the test method for the sample described in any one of [1] to [4].

[0015]

[11] A medicament for treating or preventing gynecological cancer, which is administered to a subject evaluated as having gynecological cancer by the test method described in [1]. [Advantages of the Invention]

[0016] According to the present invention, it is possible to provide a simple and highly accurate test method for a sample applicable to the evaluation of the presence or absence of gynecological cancer, the evaluation of the prognosis of a gynecological cancer patient, or the evaluation of the malignancy of gynecological cancer, a test kit usable for the test method, and a medicament for treating or preventing gynecological cancer administered to a subject identified by the test method. [Brief Description of the Drawings]

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Mode for Carrying Out the Invention

[0033] [Inspection Method of Sample (1)] The inspection method of the sample according to the first aspect can be used to evaluate the possibility that the subject has gynecological cancer. The inspection 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 derived 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 its precancerous lesions based on the concentration of the free fatty acid in the sample obtained in step (A1)

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

[0035] The stage of the gynecological cancer to be detected by the detection method of the present embodiment is not particularly limited. Note that the cancer stage described in this specification is based on the TNM classification. In the detection method of the present embodiment, it is possible to widely detect gynecological cancers from early-stage gynecological cancers (for example, Stage I, II) to advanced-stage cancers (for example, Stage III, IV). The gynecological cancer to be detected may be an early-stage gynecological malignant tumor. An early-stage gynecological malignant tumor means a gynecological malignant tumor at Stage 0, Stage I, or Stage II. An early-stage gynecological malignant tumor is preferably a malignant tumor at Stage I or Stage II. As the early-stage gynecological malignant tumor to be detected, an early-stage ovarian malignant tumor is preferred. The gynecological cancer to be detected may be a malignant tumor at Stage III or Stage IV. The gynecological cancer to be detected by the detection method of the present embodiment may be a gynecological malignant tumor having a progression degree of Stage I or higher. The gynecological malignant tumor having a progression degree of Stage I or higher includes gynecological malignant tumors at Stage I, Stage II, Stage III, and Stage IV.

[0036] In this specification, the term "precancerous lesion" means a tissue that has changed to a state in which cancer is more likely to occur compared to normal tissue. That is, a "precancerous lesion" is a tissue with a high possibility of progressing to cancer. The "precancerous lesion of gynecological cancer" means a tissue with a high possibility of progressing to "gynecological cancer". A precancerous lesion is also called an intraepithelial neoplasm.

[0037] (Step (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 is measured in a sample derived from a subject. 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 a subject is not particularly limited as long as the effects of the present invention are achieved. 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, serum, and the like.

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

[0040] In step (A1), the number of types of free fatty acids to be measured may be one or more selected from the free fatty acids in group A1, and may be 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, fourteen or more, or fifteen. The free fatty acids to be measured can be arbitrarily selected from the free fatty acids in group A1.

[0041] The free fatty acids to be measured may 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 acids to be measured may 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, or ten) 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. Alternatively, the free fatty acids to be measured may be one or more (for example, 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 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, 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 arachidic acid.

[0042] When the detection target is a gynecological 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, palmitic acid, vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, lignoceric acid, stearic acid, dihomo-γ-linolenic acid, and docosapentaenoic acid may be the measurement targets. 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 of the free fatty acids can be measured. When the detection target is a gynecological cancer, in step (A1), it is preferable to measure the concentrations of the fourteen free fatty acids.

[0043] When the gynecological cancer that can be the detection target 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 may be the measurement targets. 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 of the free fatty acids can be measured. When the gynecological cancer is ovarian cancer, in step (A1), it is preferable to measure the concentrations of the ten free fatty acids.

[0044] To enhance the detection sensitivity for early-stage gynecological malignancies (e.g., Stage I, 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 may be the measurement targets. Two or more, three or more, four or more, five or more, six or more, seven or more, or eight of the free fatty acids can be measured. When attempting to detect early-stage gynecological malignancies, in step (A1), it is preferable to measure the concentrations of the eight free fatty acids. Examples of early-stage gynecological malignancies include early-stage ovarian malignancies.

[0045] When the gynecological cancer that can be a detection target is endometrial 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 may be the measurement targets. 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 of the free fatty acids can be measured. When the gynecological cancer is endometrial cancer or cervical cancer, in step (A1), it is preferable to measure the concentrations of the twelve free fatty acids.

[0046] When the gynecological cancer that can be a detection target is endometrial 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 may be the measurement targets. Two or more, three or more, four or more, five or more, six or more, seven or more, or eight of the free fatty acids can be measured. When the gynecological cancer is endometrial cancer, in step (A1), it is preferable to measure the concentrations of the eight free fatty acids.

[0047] When the gynecological cancer that can be a detection target 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 may be the measurement targets. 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 of the free fatty acids can be measured. When the gynecological cancer is cervical cancer, in step (A1), it is preferable to measure the concentrations of the twelve free fatty acids.

[0048] When attempting to detect gynecological 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 of the above free fatty acids can be measured.

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

[0050] In step (B1), when at least one 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 its precancerous lesions.

[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 high compared to a predetermined threshold. (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 low compared to a predetermined threshold. (iii1) The concentration ratio of docosapentaenoic acid to oleic acid is low compared to a predetermined threshold.

[0052] The free fatty acids described in the above (i1) (palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, palmitic acid, and arachidonic acid) are also collectively referred to as "free fatty acids of group B(i1)". The free fatty acids described in the above (ii1) (baxenate, 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 B(ii1)". The "predetermined threshold value" in the above (i1) and (ii1) is a value set for each free fatty acid. When the free fatty acid measured in step (A1) is a free fatty acid of group B(i1), if the concentration of the free fatty acid measured in step (A1) is high compared to the predetermined threshold value set for the free fatty acid, it is evaluated that the subject is highly likely to have one or more selected from the group consisting of gynecological cancers and their precancerous lesions. When the free fatty acid measured in step (A1) is a free fatty acid of group B(ii1), if the concentration of the free fatty acid measured in step (A1) is low compared to the predetermined threshold value set for the free fatty acid, it is evaluated that the subject is highly likely to have one or more selected from the group consisting of gynecological cancers and their precancerous lesions. When two or more free fatty acids are measured in step (A1), those free fatty acids are classified into free fatty acids of group B(i1) and free fatty acids of group B(ii1), and it is confirmed whether each free fatty acid corresponds to (i1) or (ii1). The higher the ratio of the number of types of free fatty acids corresponding to (i1) or (ii1) to the number of types of measured free fatty acids, the higher the possibility that the subject has one or more selected from the group consisting of gynecological cancers and their precancerous lesions can be evaluated.

[0053] The predetermined threshold value of the concentration of each free fatty acid in the above (i1) and (ii1) may be a cut-off value obtained by acquiring the concentration of each free fatty acid from a plurality of samples derived from healthy female subjects and a plurality of samples derived from patients having one or more selected from the group consisting of gynecological cancers and their precancerous lesions, and statistically processing the concentrations of those free fatty acids.

[0054] The predetermined threshold value of the concentration of a specific free fatty acid may be set, for example, so that the positive likelihood ratio for that specific free fatty acid is maximized. The threshold value at which the positive likelihood ratio is maximized can be set, for example, by performing ROC analysis. For example, for a patient group having one or more selected from the group consisting of any number of gynecological cancers and their precancerous lesions, and a group of healthy female subjects, the serum concentration of a specific free fatty acid can be measured, and ROC analysis can be performed using those serum concentrations.

[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 respective free fatty acids in the above (i1) and (ii1) include, but are not limited to, the following.

[0056] As the threshold value of palmitoleic acid, 20 μEQ / L or more and 40 μEQ / L or less is preferable, 23 μEQ / L or more and 35 μEQ / L or less is more preferable, and 25 μEQ / L or more and 33 μEQ / L or less is still more preferable. When the gynecological cancer is ovarian malignant tumor, a specific example of the threshold value of palmitoleic acid is 29.06 μEQ / L. When attempting to detect early ovarian malignant tumor, specific examples of the threshold value are the same as those described above. When the gynecological cancer is cervical cancer, a specific example of the threshold value of palmitoleic acid is 29.12 μEQ / L. When attempting to detect including cervical cancer precancerous lesions, a specific example of the threshold value of palmitoleic acid is 36.63 μEQ / L.

[0057] As the threshold value of linoleic acid, 170 μEQ / L or more and 200 μEQ / L or less is preferable, 175 μEQ / L or more and 190 μEQ / L or less is more preferable, and 180 μEQ / L or more and 185 μEQ / L or less is still more preferable. When the gynecological cancer is ovarian malignant tumor, a specific example of the threshold value of linoleic acid is 183.1 μEQ / L. When attempting to detect early ovarian malignant tumor, specific examples of the threshold value are the same as those described above. When the gynecological cancer is cervical cancer, a specific example of the threshold value of linoleic acid is 84.63 μEQ / L. When attempting to detect including cervical precancerous lesions, a specific example of the threshold value of linoleic acid is 83.34 μEQ / L.

[0058] As the threshold value of α-linolenic acid, 20 μEQ / L or more and 40 μEQ / L or less is preferable, 23 μEQ / L or more and 35 μEQ / L or less is more preferable, and 25 μEQ / L or more and 33 μEQ / L or less is still more preferable. When the gynecological cancer is ovarian malignant tumor, a specific example of the threshold value of α-linolenic acid is 28.62 μEQ / L. When attempting to detect early ovarian malignant tumor, specific examples of the threshold value are the same as those described above. When the gynecological cancer is cervical cancer, a specific example of the threshold value of α-linolenic acid is 13.13 μEQ / L. When attempting to detect including cervical precancerous lesions, a specific example of the threshold value of α-linolenic acid is 8.334 μEQ / L.

[0059] As the threshold value of oleic acid, 300 μEQ / L or more and 35 μEQ / L or less is preferable, 310 μEQ / L or more and 340 μEQ / L or less is more preferable, and 320 μEQ / L or more and 330 μEQ / L or less is still more preferable. When the gynecological cancer is ovarian malignant tumor, a specific example of the threshold value of oleic acid is 326.0 μEQ / L or less. When attempting to detect early ovarian malignant tumor, specific examples of the threshold value are the same as those described above. When the gynecological cancer is cervical cancer, a specific example of the threshold value of oleic acid is 251.5 μEQ / L. When attempting to detect including cervical precancerous lesions, a specific example of the threshold value of oleic acid is 333.4 μEQ / L.

[0060] As the threshold value of vaccenic acid, 1.2 μEQ / L or more and 2.4 μEQ / L or less is preferable, 1.5 μEQ / L or more and 2.2 μEQ / L or less is more preferable, and 1.8 μEQ / L or more and 2.0 μEQ / L or less is even more preferable. When the gynecological cancer is ovarian malignant tumor, a specific example of the threshold value of vaccenic acid is 1.926 μEQ / L. When attempting to detect early ovarian malignant tumor, a specific example of the threshold value of vaccenic acid is 1.930 μEQ / L. When the gynecological cancer is cervical cancer, a specific example of the threshold value of vaccenic acid is 5.69 μEQ / L. When attempting to detect including cervical precancerous lesions, a specific example of the threshold value of arachidonic acid is 6.655 μEQ / L.

[0061] As the threshold value of arachidonic acid, 1.2 μEQ / L or more and 2.4 μEQ / L or less is preferable, 1.5 μEQ / L or more and 2.2 μEQ / L or less is more preferable, and 1.8 μEQ / L or more and 2.0 μEQ / L or less is even more preferable. When the gynecological cancer is ovarian malignant tumor, a specific example of the threshold value of vaccenic acid is 1.924 μEQ / L. When attempting to detect early ovarian malignant tumor, a specific example of the threshold value of arachidonic acid is 1.953 μEQ / L. When the gynecological cancer is cervical cancer, a specific example of the threshold value of arachidonic acid is 1.875 μEQ / L. When attempting to detect including cervical precancerous lesions, a specific example of the threshold value of arachidonic acid is 1.781 μEQ / L.

[0062] As the threshold value of docosahexaenoic acid, 1.3 μEQ / L or more and 2.3 μEQ / L or less is preferable, 1.5 μEQ / L or more and 2.2 μEQ / L or less is more preferable, and 1.7 μEQ / L or more and 2.03 μEQ / L or less is even more preferable. When the gynecological cancer is ovarian malignant tumor, a specific example of the threshold value of docosahexaenoic acid is 1.847 μEQ / L. When attempting to detect early ovarian malignant tumor, specific examples of the threshold value are the same as those described above. When the gynecological cancer is cervical cancer, a specific example of the threshold value of docosahexaenoic acid is 2.670 μEQ / L.

[0063] As the threshold value of adrenic acid, it 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 still more preferably 0.65 μEQ / L or more and 0.75 μEQ / L or less. When the gynecological cancer is ovarian malignant tumor, a specific example of the threshold value of adrenic acid is 0.7085 μEQ / L. When the gynecological cancer is cervical cancer, a specific example of the threshold value of adrenic acid is 0.6688 μEQ / L.

[0064] As the threshold value of nervonic acid, it 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 still more preferably 0.08 μEQ / L or more and 0.09 μEQ / L or less. When the gynecological cancer is ovarian malignant tumor, a specific example of the threshold value of nervonic acid is 0.0891 μEQ / L.

[0065] As the threshold value of lignoceric acid, it 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 still more preferably 0.5 μEQ / L or more and 0.6 μEQ / L or less. When the gynecological cancer is ovarian malignant tumor, a specific example of the threshold value of lignoceric acid is 0.5773 μEQ / L. When trying to detect early ovarian malignant tumor, specific examples of the threshold value are the same as those described above.

[0066] As the threshold value of the concentration of stearic acid, it is preferably 200 μEQ / L or more and 300 μEQ / L or less, more preferably 220 μEQ / L or more and 280 μEQ / L or less, and still more preferably 240 μEQ / L or more and 260 μEQ / L or less. Stearic acid can be preferably used for evaluation, for example, when the gynecological cancer is endometrial cancer or cervical cancer.

[0067] As the threshold value of palmitic acid, 350 μEQ / L or more and 600 μEQ / L or less is preferable, 400 μEQ / L or more and 550 μEQ / L or less is more preferable, and 430 μEQ / L or more and 500 μEQ / L or less is still more preferable. When 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 preferably used for evaluation, for example, when the gynecological cancer is endometrial cancer or cervical cancer.

[0068] As the threshold value of dihomo-γ-linolenic acid, 0.8 μEQ / L or more and 1.0 μEQ / L or less is preferable, 0.85 μEQ / L or more and 0.95 μEQ / L or less is more preferable, and 0.90 μEQ / L or more and 0.95 μEQ / L or less is still more preferable. 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] As the threshold value of docosapentaenoic acid, 0.6 μEQ / L or more and 1.5 μEQ / L or less is preferable, 0.65 μEQ / L or more and 1.2 μEQ / L or less is more preferable, and 0.7 μEQ / L or more and 1.0 μEQ / L or less is still more preferable. 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 for the concentration ratio of docosapentaenoic acid to oleic acid. Compared with the threshold value set for 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 low, it is evaluated that the subject is highly likely to have one or more selected from the group consisting of gynecological cancer and its precancerous lesions.

[0071] The predetermined threshold for the concentration ratio in the above (iii1) may be a cut-off value obtained by calculating the concentration ratio from a plurality of samples derived from healthy female subjects and a plurality of samples derived from patients having one or more selected from the group consisting of gynecological cancers and their precancerous lesions, and statistically processing these concentration ratios.

[0072] The predetermined threshold for the concentration ratio in the above (iii1) may be set, for example, so that the positive likelihood ratio for the concentration ratio is maximized. The threshold at which the positive likelihood ratio is maximized can be set by performing, for example, ROC analysis. For example, for a group of patients having one or more selected from the group consisting of gynecological cancers and their precancerous lesions and a group of healthy female subjects, the concentration ratio can be measured, and ROC analysis can be performed using these concentration ratios.

[0073] When the sample derived from the subject used in the above step (A1) is serum, examples of the predetermined threshold for the concentration ratio of docosahexaenoic acid to oleic acid include, but are not limited to, 0.004433.

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

[0075] When the gynecological cancer is ovarian cancer, the free fatty acids used for evaluation are preferably 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. By selecting the free fatty acids used for evaluation from the above 10 kinds of free fatty acids, the detection accuracy of ovarian cancer is further improved. When the gynecological cancer is ovarian cancer, for example, in step (B1), the following evaluation may be performed. When it corresponds to at least one selected from the group consisting of the following (i1)-1 and (ii1)-1, it is evaluated that the subject has a high possibility of having 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 high as compared with 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 low as compared with a predetermined threshold value.

[0076] In order to enhance the detection sensitivity of early gynecological malignancies (e.g., Stage I, II), 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, vaccenic acid, arachidic acid, docosahexaenoic acid, and lignoceric acid. By selecting the free fatty acids used for evaluation from the above eight free fatty acids, the detection accuracy of early gynecological malignancies (particularly, ovarian malignancies at Stage I or II) is further improved. When attempting to detect early gynecological malignancies, for example, in step (B1), the following evaluations may be performed. When corresponding to at least one selected from the group consisting of the following (i1)-2 and (ii1)-2, it is evaluated that the subject is highly likely to have a gynecological cancer (e.g., ovarian malignancy) with a progression degree of Stage I or higher; (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 high as compared with 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 low as compared with a predetermined threshold value.

[0077] When the gynecological cancer is endometrial cancer or cervical cancer, the free fatty acids used for evaluation are preferably 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. By selecting the free fatty acids used for evaluation from the above 12 free fatty acids, the detection accuracy of endometrial cancer or cervical cancer can be further improved. When the gynecological cancer is endometrial cancer or cervical cancer, for example, in step (B1), the following evaluations may be performed. When it corresponds to at least one selected from the group consisting of the following (i1)-3 and (ii1)-3, 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 high compared to a predetermined threshold; (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 low compared to a predetermined threshold.

[0078] When the gynecological cancer is endometrial cancer, the free fatty acids used for evaluation are preferably 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. By selecting the free fatty acids used for evaluation from the above 8 free fatty acids, the detection accuracy of endometrial cancer can be further improved. When the gynecological cancer is endometrial cancer, for example, in step (B1), the following evaluations may be performed. When it corresponds to at least one selected from the group consisting of the following (i1)-4 and (ii1)-4, it is evaluated that the subject is highly likely to have ovarian cancer; (i1) - 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 high compared to 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, and adrenic acid in the sample is low compared to a predetermined threshold value.

[0079] When the gynecological cancer is cervical cancer, the free fatty acid used for evaluation is preferably 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. By selecting the free fatty acid used for evaluation from the above 12 free fatty acids, the detection accuracy of cervical cancer is further improved. When the gynecological cancer is cervical cancer, for example, in step (B1), the following evaluation may be performed. When it corresponds to at least one selected from the group consisting of the following (i1)-5 and (ii1)-5, it is evaluated that the subject has a high possibility of having 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 high compared to 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 low compared to a predetermined threshold value.

[0080] When attempting to detect gynecological precancerous lesions (e.g., cervical cancer precancerous lesions), the free fatty acids used for evaluation are preferably one or more free fatty acids selected from the group consisting of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, arachidonic acid, and eicosanoic acid. When the detection target is gynecological cancer and its precancerous lesions, for example, in step (B1), the following evaluations may be performed. When corresponding to at least one selected from the group consisting of the following (i1)-6 and (ii1)-6, it is evaluated that the subject is highly likely to have a gynecological precancerous lesion and / or 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 high compared to a predetermined threshold; (ii1)-6 The concentration of arachidonic acid free fatty acid in the sample is low compared to a predetermined threshold.

[0081] In step (B1), the step of determining whether it corresponds to one or more selected from the group consisting of the above (i1) and (ii1) is the step of determining that the subject is highly likely to have gynecological cancer when the total value of the scores of each free fatty acid (hereinafter referred to as "FFA score") set in the following (i-1) and (ii-1) is higher than a predetermined threshold (hereinafter also referred to as "step (B1-1)").

[0082] (i-1) When the free fatty acid is a free fatty acid in group B(i1), when the concentration of the free fatty acid in the sample is high compared to a predetermined threshold, " +1" is set as the score of the free fatty acid. When the concentration of the free fatty acid in the sample is low compared to a predetermined threshold, "0" is set as the score of the free fatty acid. (ii-1) When the free fatty acid is a free fatty acid in group B(ii1), when the concentration of the free fatty acid in the sample is low compared to a predetermined threshold, " +1" is set as the score of the free fatty acid. When the concentration of the free fatty acid in the sample is high compared to a predetermined threshold, "0" is set as the score of the free fatty acid. For each free fatty acid used in (i-1) and (ii-1), examples of the predetermined threshold values are the same as those listed as the predetermined threshold values in (i1) and (ii1). Calculate the total value of the scores of each free fatty acid set by (i-1) and (ii-1) and use it as the FFA score.

[0083] The predetermined threshold value for the FFA score may be a cut-off value obtained by calculating the FFA scores for a plurality of samples derived from healthy female subjects and a plurality of samples derived from patients having one or more selected from the group consisting of gynecological cancers and their precancerous lesions, and statistically processing those FFA scores.

[0084] The predetermined threshold value for the FFA score may be set, for example, so that the positive likelihood ratio for the FFA score is maximized. The threshold value that maximizes the positive likelihood ratio can be set, for example, by performing ROC analysis. For example, for a group of patients having one or more selected from the group consisting of gynecological cancers and their precancerous lesions and a group of healthy female subjects, the serum concentration of a specific free fatty acid can be measured, and ROC analysis can be performed using those serum concentrations.

[0085] The predetermined threshold value for the FFA score depends on the number of types of free fatty acids used in the calculation of the FFA score. For example, it may be 1 or more and 3 or less, preferably more than 1 and less than 3, more preferably more than 1 and 2 or less, and even more preferably more than 1 and less than 2. Examples of the number of types of free fatty acids used in the calculation of the FFA score 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, 14 or more, 15 of the free fatty acids in the above A1 group.

[0086] When the gynecological cancer is ovarian cancer, as the free fatty acids used for calculating the FFA score, one or more 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 are preferable. The free fatty acids used for calculating the FFA score may 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 above free fatty acids. When the gynecological cancer is ovarian cancer, it is preferable to use the 10 free fatty acids for calculating the FFA score. When using the 10 free fatty acids, examples of the predetermined threshold value of the FFA score include the same as above, and for example, it can be set to more than 1 and less than 2 (for example, 1.5).

[0087] To enhance the detection sensitivity of early gynecological malignancies (e.g., Stage I, II), as the free fatty acids used for calculating the FFA score, 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 may be used. The free fatty acids used for calculating the FFA score may be 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, or 8 of the above free fatty acids. From the viewpoint of the detection sensitivity of early gynecological malignancies, it is preferable to use the 8 free fatty acids for calculating the FFA score. When using the 8 free fatty acids, examples of the predetermined threshold value of the FFA score include the same as above, and for example, it can be set to more than 1 and less than 2 (for example, 1.5).

[0088] When the gynecological cancer is endometrial cancer or cervical cancer, as the free fatty acids used for calculating the FFA score, 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, adrenic acid and docosapentaenoic acid are preferable. The free fatty acids used for calculating the FFA score may 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 above-mentioned free fatty acids. The number of types of free fatty acids used for calculating the FFA score and the predetermined threshold value of the FFA score can be set as appropriate.

[0089] When the gynecological cancer is endometrial cancer, as the free fatty acids used for calculating the FFA score, 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 are preferable. The free fatty acids used for calculating the FFA score may be 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, or 8 of the above-mentioned free fatty acids. The number of types of free fatty acids used for calculating the FFA score and the predetermined threshold value of the FFA score can be set as appropriate.

[0090] When the gynecological cancer is cervical cancer, as the free fatty acids used for calculating the FFA score, 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, adrenic acid and docosapentaenoic acid are preferable. The free fatty acids used for calculating the FFA score may 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 above-mentioned free fatty acids. The number of types of free fatty acids used for calculating the FFA score and the predetermined threshold value of the FFA score can be set as appropriate.

[0091] In the inspection method according to the first aspect, when using the FFA score, specific examples of steps (A1) and (B1) are as follows. (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 the subject are measured. In step (B1), when the FFA score calculated from 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 highly likely to have gynecological cancer. As the threshold of the FFA score calculated from the above 10 free fatty acids, 1 or more and 3 or less is preferable, 1.2 or more and 2.5 or less is more preferable, 1.3 or more and 2 or less is still more preferable, and 1.3 or more and 1.9 or less is particularly preferable.

[0092] (Example 2) In step (A1), the concentrations of 8 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 the subject are measured. In step (B1), when the FFA score calculated from 8 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 highly likely to have gynecological cancer. As the threshold of the FFA score of the above 8 free fatty acids, 0.5 or more and 2.5 or less is preferable, 1 or more and 2 or less is more preferable, and 1.1 or more and 1.9 or less is still more preferable.

[0093] By using the FFA score, it is possible to more accurately evaluate the possibility that the subject has one or more selected from the group consisting of gynecological cancers and their precancerous lesions. By using the FFA score, it is possible to accurately detect 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). These tumors had a high proportion of false negatives and low sensitivity with conventional tumor markers such as CA125. However, by using the FFA score, these tumors can also be detected with high sensitivity. For example, regarding the early gynecological malignancies, by using the FFA score, gynecological malignancies at Stage I or Stage II can also be detected with high sensitivity.

[0094] In step (B1), a subject evaluated as having a high possibility of having one or more selected from the group consisting of gynecological cancers and their precancerous lesions is likely to have one or more selected from the group consisting of any cancer among gynecological cancers and their precancerous lesions. Therefore, in order to identify the type of one or more selected from the group consisting of the gynecological cancers and their precancerous lesions that the subject has, further examinations may be performed by computed tomography (CT) examination, magnetic resonance imaging (MRI), positron emission tomography (PET) examination, ultrasonic examination, etc. Furthermore, for a definitive diagnosis, a pathological examination (cytological examination, histological examination) may be performed.

[0095] The method for examining a sample according to the first aspect may further include the following step (C1). Step (C1): Evaluate, based on the concentration of the free fatty acid in the sample obtained in step (A1), which of the gynecological cancer and the gynecological precancerous lesion the subject determined in step (B1) to have a high possibility of having one or more selected from the group consisting of gynecological cancers and their precancerous lesions has.

[0096] The subject in step (C1) is the subject determined in step (B1) to have a high possibility of having one or more selected from the group consisting of gynecological cancers and their precancerous lesions.

[0097] (Step (C1)) In step (B1), based on the concentration of free fatty acids in the sample obtained in step (A1), it is evaluated whether the subject has either gynecological cancer or a gynecological pre-cancerous lesion.

[0098] The gynecological cancer to be determined is preferably cervical cancer. The gynecological pre-cancerous lesion to be determined is preferably a cervical pre-cancerous lesion.

[0099] In step (C1), when the following (Ci) is satisfied, it is evaluated that the subject has a high possibility of having 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 low as compared with a predetermined threshold value.

[0100] Hereinafter, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid are also collectively referred to as "free fatty acids of group C1".

[0101] In step (C1), the number of types of free fatty acids to be determined may be one or more selected from the free fatty acids of group C1, may be two or more, may be three or more, or may be four. The free fatty acids to be determined can be arbitrarily selected from among the free fatty acids of group C1.

[0102] 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) may be a cut-off value obtained by acquiring the concentration of each free fatty acid from a plurality of samples derived from patients with gynecological cancer and a plurality of samples derived from patients with gynecological pre-cancerous lesions and performing statistical processing on the concentrations of those free fatty acids.

[0103] A predetermined threshold value for the concentration of a specific free fatty acid may be set, for example, such that the positive likelihood ratio for that specific free fatty acid is maximized. The threshold value at which the positive likelihood ratio is maximized can be set, for example, by performing ROC analysis. For example, for a group of patients with gynecological cancer and a group of patients with gynecological precancerous lesions, the serum concentration of a specific free fatty acid can be measured, and ROC analysis can be performed using those serum concentrations.

[0104] Examples of the threshold values for the concentrations of each free fatty acid in the above (Ci) include, but are not limited to, the following.

[0105] As the threshold value of dihomo-γ-linolenic acid, it 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 even more preferably 0.95 μEQ / L or more and 1.05 μEQ / L or less. When the gynecological cancer is cervical cancer and the gynecological precancerous lesion is a cervical precancerous lesion, a specific example of the threshold value of dihomo-γ-linolenic acid is 1.012 μEQ / L.

[0106] As the threshold value of docosahexaenoic acid, it 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 even 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 precancerous lesion is a cervical precancerous lesion, a specific example of the threshold value of docosahexaenoic acid is 3.507 μEQ / L.

[0107] As the threshold value of arachidonic acid, it 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 even 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 precancerous lesion is a cervical precancerous lesion, a specific example of the threshold value of docosahexaenoic acid is 0.312 μEQ / L.

[0108] As the threshold value of docosahexaenoic acid, it 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 precancerous lesion is a cervical precancerous lesion, a specific example of the threshold value of docosahexaenoic acid is 0.7231 μEQ / L.

[0109] Step (C1) may be a step of calculating an FFA score and evaluating that the subject is likely to have a gynecological cancer when the FFA score is higher than a predetermined threshold value. The FFA score is calculated by the same method as the method described in the above first aspect. 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 the above (i-1) and (ii-1).

[0110] The predetermined threshold value for the FFA score may be a cut-off value obtained by calculating the FFA score for a plurality of samples derived from gynecological cancer patients and a plurality of samples derived from patients with precancerous lesions and statistically processing those FFA scores.

[0111] The predetermined threshold value for the FFA score depends on the number of types of free fatty acids used in the calculation of the FFA score. For example, when the number of types of free fatty acids is 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. Examples of the number of types of free fatty acids used in the calculation of the FFA score include 3 or more and 4 types among the free fatty acids in the above Group A5.

[0112] For a subject confirmed to have one or more selected from the group consisting of gynecological cancers and their precancerous lesions by the above-described diagnosis, one or more treatments selected from the group consisting of gynecological cancers and their precancerous lesions can be performed. In one embodiment, the present invention provides a method for treating one or more selected from the group consisting of gynecological cancers and their precancerous lesions, comprising: (a1) identifying a subject highly likely to have one or more selected from the group consisting of gynecological cancers and their precancerous lesions by the method for examining a sample according to the first aspect as described above; (b1) confirming that the subject has one or more selected from the group consisting of gynecological cancers and their precancerous lesions; and (c1) performing one or more treatments selected from the group consisting of gynecological cancers and their precancerous lesions on the subject confirmed to have one or more selected from the group consisting of gynecological cancers and their precancerous lesions. In the step (a1), the method for examining a sample according to the first aspect is carried out as described above to identify a subject highly likely to have one or more selected from the group consisting of gynecological cancers and their precancerous lesions. In the step (b1), further examination is performed on the subject identified in the step (a1) to confirm that the subject has one or more selected from the group consisting of gynecological cancers and their precancerous lesions. Examples of the one or more examinations selected from the group consisting of gynecological cancers and their precancerous lesions include the above-described examinations. In the step (c1), for a subject confirmed to have one or more selected from the group consisting of gynecological cancer and its precancerous lesions in the step (b1), one or more treatments selected from the group consisting of gynecological cancer and its precancerous lesions can be performed. The treatment method of gynecological cancer is not particularly limited, and examples include surgical treatment, chemotherapy, and radiotherapy. For example, when the gynecological cancer is early-stage gynecological cancer (e.g., Stage I or Stage II), as the main treatment method, for example, surgical treatment can be selected. In the case of early-stage gynecological cancer, for example, if the gynecological cancer can be completely removed by surgical treatment or the like, the gynecological cancer can be cured. In the case of advanced gynecological cancer (e.g., Stage III or Stage IV), a combination of surgical treatment, chemotherapy (such as administration of anticancer drugs), and radiotherapy may be used for treatment. For example, chemotherapy and / or radiotherapy may be performed before surgical treatment to shrink the tumor. Alternatively, after surgical treatment, chemotherapy and / or radiotherapy may be performed to shrink the tumor remaining after surgical treatment. The anticancer drugs used in chemotherapy include the anticancer drugs described later in the section of [Medicine]. As a treatment method for gynecological cancer precancerous lesions, for example, vaginal surgeries such as cervical conization and cervical laser evaporation may be performed.

[0113] According to the method for examining a sample according to the first aspect described above, it is possible to simply, with high specificity and high sensitivity, determine the possibility that a subject has one or more selected from the group consisting of gynecological cancer and its precancerous lesions. Diagnosis using CA125 as a marker has low detection sensitivity for early ovarian malignancies and ovarian clear cell malignancies. The examination method according to the first aspect can detect with high specificity and high sensitivity even when a subject has an early ovarian malignancy or an ovarian clear cell malignancy.

[0114] Advanced gynecological cancers often have a high risk of recurrence even after treatment and often have a poor prognosis. On the other hand, if gynecological cancer is detected at an early stage before it progresses, for example, through treatment such as tumor resection, it is possible to completely cure the gynecological cancer. Alternatively, the prognosis can be improved. According to the method for examining a sample according to the first aspect, since it is possible to detect early-stage gynecological cancer, it is possible to increase the probability of completely curing gynecological cancer through treatment.

[0115] Due to the risk of metastasis in cervical cancer, invasive surgical treatments such as radical / subtotal hysterectomy and lymph node dissection, or treatments such as radiation therapy are performed. Therefore, in young people in their 30s and 40s, which is the peak incidence, fertility may be lost. On the other hand, cervical cancer precancerous lesions are tumors limited to the epithelium without invasion, so there is no possibility of metastasis. Therefore, less invasive treatments such as cervical conization and cervical laser evaporation are performed, and fertility is also preserved. Therefore, detecting cervical cancer precancerous lesions is a great benefit to the subject.

[0116] Step (B1) may be a step of identifying a sample derived from a subject who is likely to have one or more selected from the group consisting of the gynecological cancer and its precancerous lesions when it corresponds to at least one selected from the group consisting of (i1) and (ii1). [[ID=X]]

[0117] Step (C1) may be a step of identifying a sample derived from a subject who is likely to have the gynecological cancer when it corresponds to (Ci).

[0118] [Method for Examining a Sample (2)] The method for examining a sample according to the second aspect is a method for evaluating the possibility that a subject with gynecological cancer will have a poor prognosis. The examination method includes step (A2) and step (B2). Step (A2): 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): Evaluating the likelihood of poor prognosis based on the concentration of free fatty acids in the sample obtained in step (A2)

[0119] In the method for examining a sample according to the second aspect, the subject from which the sample to be examined is derived is a human female having a gynecological cancer. For example, the subject is a human female who has been previously diagnosed with a gynecological cancer. The subject may be before the start of treatment for a gynecological cancer, during treatment for a gynecological cancer, or may have received treatment for a gynecological cancer in the past. Alternatively, it may be a subject determined to have a gynecological cancer by the examination method according to the first aspect. Examples of the gynecological cancer include the cancers described above in the first aspect. The gynecological cancer is preferably a gynecological malignancy, more preferably one or more selected from the group consisting of ovarian malignancy, uterine body malignancy, and cervical malignancy, and even more preferably ovarian malignancy.

[0120] 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".

[0121] Step (A2) can be performed in the same manner as step (A1) in the first aspect, except that a sample derived from a subject having a gynecological cancer is used.

[0122] In step (A2), the number of types of free fatty acids to be measured may be one or more selected from the free fatty acids in group A2, and may be 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. The free fatty acids to be measured can be arbitrarily selected from the free fatty acids in group A2.

[0123] 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 may be used as the measurement target. 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 of the free fatty acids can be measured. When the gynecological cancer is ovarian cancer, in step (A2), it is preferable to measure the concentrations of the ten free fatty acids.

[0124] When the gynecological cancer is early-stage gynecological malignancy (for example, 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 lignoceric acid may be used as the measurement target. Two or more, three or more, four or more, five or more, six or more, seven or more, or eight of the free fatty acids can be measured. When attempting to detect early-stage gynecological malignancy, in step (A2), it is preferable to measure the concentrations of the eight free fatty acids. Examples of early-stage gynecological malignancy include early-stage ovarian malignancy.

[0125] When the gynecological cancer is endometrial 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, adrenic acid, and docosapentaenoic acid may be the measurement targets. 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 of the free fatty acids can be measured. When the gynecological cancer is endometrial cancer or cervical cancer, in step (A2), it is preferable to measure the concentrations of the twelve free fatty acids.

[0126] When the gynecological cancer is endometrial 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 may be the measurement targets. Two or more, three or more, four or more, five or more, six or more, seven or more, or eight of the free fatty acids can be measured. When the gynecological cancer is endometrial cancer, in step (A2), it is preferable to measure the concentrations of the eight free fatty acids.

[0127] 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 may be the measurement targets. 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 of the free fatty acids can be measured. When the detection target is cervical cancer, in step (A2), it is preferable to measure the concentrations of the twelve free fatty acids.

[0128] In step (B2), when it corresponds to at least one selected from the group consisting of the following (i2) and (ii2), it is evaluated that the subject has a high possibility of 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 high compared to 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 low compared to a predetermined threshold value.

[0129] The "predetermined threshold value" in the above (i2) and (ii2) is a value set for each free fatty acid. The predetermined threshold value of the concentration of each free fatty acid may be a cut-off value obtained by obtaining the concentration of each free fatty acid from a plurality of samples derived from gynecological patients with good prognosis and a plurality of samples derived from gynecological cancer patients with poor prognosis, and statistically processing the concentrations of those free fatty acids.

[0130] Step (B2) may be a step of calculating an FFA score and evaluating that the subject has a high possibility of poor prognosis of gynecological cancer when the FFA score is higher than a predetermined threshold value. The FFA score is calculated by the same method as the method described in the first aspect. 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 the above (i-1) and (ii-1).

[0131] In step (B2), the predetermined threshold value set in the same manner as the above (i-1) and (ii-1) may be obtained as described above, or the same threshold value as the inspection method according to the first aspect may be used. A predetermined threshold value for the FFA score may be a cut-off value obtained by calculating the FFA scores for a plurality of samples derived from gynecological cancer patients with a favorable prognosis and a plurality of samples derived from gynecological cancer patients with an unfavorable prognosis, and statistically processing those FFA scores. Alternatively, the FFA scores may be calculated for a plurality of samples derived from gynecological cancer patients, and the median of those FFA scores may be used as the threshold value for the FFA score.

[0132] A predetermined threshold value for the FFA score also depends on the number of types of free fatty acids used in the calculation of the FFA score. For example, when the number of types of free fatty acids is n, examples include 0.4n or more and 0.8n or less, 0.5n or more and 0.7n or less, or 0.5n or more and 0.6n or less. Examples of the number of types of free fatty acids used in the calculation of the FFA score include 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more of the free fatty acids in the above Group A2. "0.4n" means the value obtained by multiplying 0.4 by the number of types n. The same applies to other numerical values. For example, when the number of types of free fatty acids is 5, "0.4n or more and 0.8n or less" means "2 or more and 4 or less".

[0133] When the gynecological cancer is ovarian cancer, the free fatty acid used in the calculation of the FFA score is preferably at least one 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 free fatty acid used in the calculation of the FFA score may 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 above free fatty acids. When the gynecological cancer is ovarian cancer, it is preferable to use the above 10 free fatty acids in the calculation of the FFA score. A predetermined threshold value for the FFA score is the same as the predetermined threshold value for FFA described above in step (B2). When using the above 10 free fatty acids, examples of the predetermined threshold value for the FFA score include more than 4 and less than 6.

[0134] When gynecological cancer is an early-stage gynecological malignant tumor, as the free fatty acids used for calculating the FFA score, 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 are preferable. The free fatty acids used for calculating the FFA score may be 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, or 8 of the above free fatty acids. When attempting to detect early-stage gynecological malignant tumors, it is preferable to use the above 8 free fatty acids for calculating the FFA score. The predetermined threshold value for the FFA score is the same as the predetermined threshold value for FFA described above in step (B2). When using the above 8 free fatty acids, examples of the predetermined threshold value for the FFA score include more than 3 and less than 5.

[0135] When gynecological cancer is endometrial cancer or cervical cancer, as the free fatty acids used for calculating the FFA score, 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, adrenic acid, and docosapentaenoic acid are preferable. The free fatty acids used for calculating the FFA score may 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 above free fatty acids. The number of types of free fatty acids used for calculating the FFA score and the predetermined threshold value of the FFA score can be set as appropriate. When gynecological cancer is endometrial cancer or cervical cancer, it is preferable to use the above 12 free fatty acids for calculating the FFA score. The predetermined threshold value for the FFA score is the same as the predetermined threshold value for FFA described above in step (B2).

[0136] When the gynecological cancer is endometrial cancer, as the free fatty acids used for calculating the FFA score, 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 are preferable. The free fatty acids used for calculating the FFA score may be 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, or 8 of the above free fatty acids. The number of types of free fatty acids used for calculating the FFA score and the predetermined threshold value of the FFA score can be set as appropriate. When the gynecological cancer is endometrial cancer, it is preferable to use the above 8 free fatty acids for calculating the FFA score. The predetermined threshold value for the FFA score is the same as the predetermined threshold value for FFA described above in step (B2).

[0137] When the gynecological cancer is cervical cancer, as the free fatty acids used for calculating the FFA score, 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, adrenic acid, and docosapentaenoic acid are preferable. The free fatty acids used for calculating the FFA score may 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 above free fatty acids. The number of types of free fatty acids used for calculating the FFA score and the predetermined threshold value of the FFA score can be set as appropriate. When the gynecological cancer is cervical cancer, it is preferable to use the above 10 free fatty acids for calculating the FFA score. The predetermined threshold value for the FFA score is the same as the predetermined threshold value for FFA described above in step (B2).

[0138] In the inspection method according to the second aspect, when using the FFA score, specific examples of step (A2) and step (B2) are as follows. 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 highly likely to have a poor prognosis. The predetermined threshold value for the FFA score may be a cut-off value obtained by calculating the FFA scores for 10 free fatty acids derived from gynecological cancer patients with a good prognosis and 10 free fatty acids derived from gynecological cancer patients with a poor prognosis, and statistically processing these FFA scores. Alternatively, the FFA scores may be calculated for a plurality of samples derived from gynecological cancer patients, and the median of these FFA scores may be used as the threshold value for the FFA score.

[0139] According to the method for examining a sample according to the second aspect described above, it is possible to predict whether a subject will have a poor prognosis. Therefore, referring to this test result makes it easier to establish an appropriate treatment plan. Also, it becomes easier to predict recurrence after treatment. When the score is higher than a predetermined threshold value, it is evaluated that the subject is highly likely to have a poor prognosis.

[0140] [[ID= twelve]]For example, for a subject evaluated as being highly likely to have a poor prognosis, measures such as changing the treatment method may be taken. In one embodiment, the present invention provides a method for treating gynecological cancer, including: (a2) a step of identifying a subject highly likely to have a poor prognosis by the method for examining a sample according to the second aspect, and (b2) a step of changing the treatment method for gynecological cancer for the subject. In step (a2), the method for examining a sample according to the second aspect is carried out as described above to identify a subject highly likely to have a poor prognosis. In the step (b2), in the subject identified in step (a2), the treatment method for gynecological cancer can be reexamined and the treatment method for gynecological cancer can be changed. The subject identified in step (a2) is likely to have a poor prognosis even if the previous treatment is continued. Therefore, by reexamining the treatment method and changing the treatment method, it is possible to explore the possibility of improving the prognosis. Examples of changes in the treatment method include changes in the type of anticancer agent.

[0141] Step (B2) may be a step of identifying a sample derived from a subject with a high likelihood of a poor prognosis when it corresponds to at least one selected from the group consisting of the above (i2) and (ii2).

[0142] [Inspection method of sample (3)] The inspection method of the sample according to the third aspect 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 step (A3) and 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)

[0143] In the inspection method of the sample according to the third aspect, the subject from which the sample used for the inspection is derived is a human female having gynecological cancer. Examples of the gynecological cancer possessed by the subject include those described above in the first aspect. For example, the subject is a human female who has been previously diagnosed with having gynecological cancer. Alternatively, it may be a subject determined to have gynecological cancer by the inspection method according to the first aspect described above.

[0144] As the gynecological malignant tumor, it is preferably at least one selected from the group consisting of ovarian malignant tumor, uterine body malignant tumor, and cervical malignant tumor, and ovarian malignant tumor is more preferable. As the gynecological borderline malignant tumor, it 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 ovarian borderline malignant tumor is preferable. In the evaluation in the inspection method according to the third aspect, for example, it may be to evaluate whether the ovarian cancer possessed by the subject is an ovarian malignant tumor or an ovarian borderline malignant tumor. It may be to evaluate whether the uterine body cancer possessed by the subject is a uterine body malignant tumor or a uterine body borderline malignant tumor. It may be to evaluate whether the cervical cancer possessed by the subject is a cervical malignant tumor or a cervical borderline malignant tumor.

[0145] 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 collectively also referred to as "free fatty acids of group A3".

[0146] Step (A3) can be carried out in the same manner as step (A1) in the first aspect, except that a sample derived from a subject having a gynecological cancer is used.

[0147] In step (A3), the number of types of free fatty acids to be measured may be one or more selected from the free fatty acids of group A3, and may be 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. The free fatty acids to be measured can be arbitrarily selected from the free fatty acids of group A3.

[0148] 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 may be the measurement targets. 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 of the free fatty acids can be measured. When the gynecological cancer is ovarian cancer, in step (A3), it is preferable to measure the concentrations of the ten free fatty acids.

[0149] When the gynecological cancer is endometrial 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, adrenic acid, and docosapentaenoic acid may be the measurement targets. 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 of the free fatty acids can be measured. When the gynecological cancer is endometrial cancer or cervical cancer, in step (A3), it is preferable to measure the concentrations of the twelve free fatty acids.

[0150] When the gynecological cancer is endometrial 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 may be the measurement targets. Two or more, three or more, four or more, five or more, six or more, seven or more, or eight of the free fatty acids can be measured. When the gynecological cancer is endometrial cancer, in step (A3), it is preferable to measure the concentrations of the eight free fatty acids.

[0151] 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 may be the measurement targets. 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 of the free fatty acids can be measured. When the detection target is cervical cancer, in step (A3), it is preferable to measure the concentrations of the twelve free fatty acids.

[0152] In step (B3), when at least one of the following (i3) and (ii3) is satisfied, it is evaluated that the gynecological cancer of 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 high compared to 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 low compared to a predetermined threshold value.

[0153] The "predetermined threshold value" in the above (i3) and (ii3) is a value set for each free fatty acid. The predetermined threshold value of the concentration of each free fatty acid may be a cut-off value obtained by acquiring the concentrations of each free fatty acid from a plurality of samples derived from patients with gynecological malignant tumors and a plurality of samples derived from patients with borderline gynecological malignant tumors, and statistically processing the concentrations of those free fatty acids.

[0154] Step (B3) may be a step of calculating an FFA score and, when the FFA score is higher than a predetermined threshold, 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 the method described in the first aspect. 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 the above (i-1) and (ii-1).

[0155] In step (B3), the predetermined threshold set in the same manner as the above (i-1) and (ii-1) may be obtained as described above, or the same threshold as the inspection method according to the first aspect may be used. The predetermined threshold for the FFA score may be a cut-off 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 borderline gynecological malignancies and statistically processing those FFA scores.

[0156] The predetermined threshold for the FFA score depends on the number of types of free fatty acids used in the calculation of the FFA score. For example, when the number of types of free fatty acids is 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. Examples of the number of types of free fatty acids used in the calculation of the FFA score include 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more of the free fatty acids in the above group A3.

[0157] When the gynecological cancer is ovarian cancer, as the free fatty acids used for calculating the FFA score, one or more 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 are preferable. The free fatty acids used for calculating the FFA score may 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 above free fatty acids. When the gynecological cancer is ovarian cancer, it is preferable to use the 10 free fatty acids for calculating the FFA score. When using the 10 free fatty acids, examples of the predetermined threshold value of the FFA score include more than 4 and less than 6.

[0158] When the gynecological cancer is endometrial cancer or cervical cancer, as the free fatty acids used for calculating the FFA score, 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, adrenic acid and docosapentaenoic acid are preferable. The free fatty acids used for calculating the FFA score may 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 above free fatty acids. When the gynecological cancer is endometrial cancer or cervical cancer, it is preferable to use the 12 free fatty acids for calculating the FFA score. When using the 12 free fatty acids, examples of the predetermined threshold value of the FFA score include the same as those described above.

[0159] When the gynecological cancer is endometrial cancer, as the free fatty acids used for calculating the FFA score, 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 are preferable. The free fatty acids used for calculating the FFA score may be 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, or 8 of the above free fatty acids. When the gynecological cancer is endometrial cancer, it is preferable to use the above 8 free fatty acids for calculating the FFA score. When using the above 8 free fatty acids, examples of the predetermined threshold value of the FFA score are the same as those described above.

[0160] When the gynecological cancer is cervical cancer, as the free fatty acids used for calculating the FFA score, 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 are preferable. The free fatty acids used for calculating the FFA score may 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 above free fatty acids. When the gynecological cancer is cervical cancer, it is preferable to use the above 12 free fatty acids for calculating the FFA score. When using the above 12 free fatty acids, examples of the predetermined threshold value of the FFA score are the same as those described above.

[0161] In the inspection method according to the third aspect, when using the FFA score, specific examples of step (A3) and step (B3) are as follows. 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 ten 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 determined that the gynecological cancer possessed by the subject is likely to be a malignant tumor. In this case, as the threshold value of the total score, it is preferably 4 or more and 6 or less, more preferably 4.5 or more and 5 or less.

[0162] According to the method for examining a sample according to 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 borderline gynecological malignant tumor. A borderline malignant tumor is a tumor having intermediate properties between a malignant tumor and a benign tumor. Since an ovarian borderline malignant tumor does not metastasize hematogenously or lymphatically like a malignant tumor, its prognosis is not as poor as that of a malignant tumor. That is, an ovarian borderline malignant tumor has the clinical feature that although the lesion may rarely spread intraperitoneally, there is no metastasis. In addition, the growth rate of the tumor generally differs between a borderline malignant tumor and a malignant tumor. A malignant tumor grows rapidly and requires surgical treatment as soon as possible because of seeding and metastasis. On the other hand, a borderline malignant tumor has slow tumor growth and rarely requires urgent surgery. Due to the above differences in characteristics, non-invasively identifying whether a tumor is a borderline malignant tumor or a malignant tumor by a blood test before treatment (at the time of diagnosis) is extremely useful for determining the treatment policy (waiting time until surgery, surgical procedure, degree of surgical invasion, etc.). Currently, intraoperative pathological diagnosis is used to identify which tumor it is, but by the method for examining a sample according to the third aspect, it is possible to non-invasively evaluate before surgery whether the gynecological cancer possessed by the subject is a borderline malignant tumor or a malignant tumor.

[0163] In one embodiment, the present invention provides a method for treating gynecological cancer, comprising: (a3) identifying a subject highly likely to have a borderline gynecological malignancy or a subject highly likely to have a gynecological malignancy by the method for examining a sample according to the third aspect; and (b3) treating the subject highly likely to have a borderline gynecological malignancy for borderline gynecological malignancy, and treating the subject highly likely to have a gynecological malignancy for gynecological malignancy. In the step (a3), the method for examining a sample according to the third aspect is carried out as described above to identify a subject highly likely to have a borderline gynecological malignancy or a subject highly likely to have a gynecological malignancy. In the step (b3), in the subject identified in the step (a3), when the subject is highly likely to have a borderline gynecological malignancy, treatment for borderline gynecological malignancy is carried out. Treatment for borderline gynecological malignancy includes surgical treatment, preferably removal of intraperitoneal organs (i.e., uterus, ovaries, fallopian tubes and omentum), and more preferably removal of only intraperitoneal organs. Also, when the subject is highly likely to have a gynecological malignancy, treatment for gynecological malignancy is carried out. Treatment for gynecological malignancy includes surgical treatment, preferably removal of intraperitoneal organs and lymph node dissection.

[0164] The step (B3) may be a step of identifying a sample derived from a subject in which there is a high possibility that the gynecological cancer is a malignant tumor when it corresponds to at least one selected from the group consisting of the above (i3) and (ii3).

[0165] [Kit] The kit for examining gynecological cancer and its precancerous lesions according to the fourth aspect 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, 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 this aspect is used for at least one purpose selected from the group consisting of the following (a) to (d). (a) Evaluation of the possibility that a subject has one or more selected from the group consisting of gynecological cancers and their precancerous lesions; (b) Evaluation of the possibility that a subject with a gynecological cancer has a poor prognosis; (c) Evaluation of whether the gynecological cancer that a subject has is a malignant tumor or a borderline malignant tumor; (d) Evaluation of which of gynecological cancers and gynecological precancerous lesions a subject highly likely to have one or more selected from the group consisting of gynecological cancers and gynecological precancerous lesions has.

[0166] The subject in the above (a) is a human female. Examples of the gynecological cancers that the subject may have include those described above in the first aspect. As the gynecological cancers that the subject may have, gynecological malignant tumors are preferred. As the gynecological malignant tumors, one or more selected from the group consisting of ovarian malignant tumors, uterine body malignant tumors, and cervical malignant tumors are preferred, and ovarian malignant tumors are more preferred. As the gynecological borderline malignant tumors, one or more selected from the group consisting of ovarian borderline malignant tumors, uterine body borderline malignant tumors, and cervical borderline malignant tumors are preferred, and ovarian borderline malignant tumors are more preferred. Examples of the gynecological precancerous lesions that the subject may have include those described above in the first aspect.

[0167] The subject in the above (b) is a human female with a gynecological cancer. Examples of the gynecological cancers include those cancers described above in the first aspect. As the gynecological cancers, gynecological malignant tumors are preferred, one or more selected from the group consisting of ovarian malignant tumors, uterine body malignant tumors, and cervical malignant tumors are more preferred, and ovarian malignant tumors are even more preferred.

[0168] The subject in the above (c) is a human female having gynecological cancer. Examples of the gynecological cancer include the cancers described above in the first aspect. As the gynecological malignant tumor, it 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. As the gynecological borderline malignant tumor, it 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 ovarian borderline malignant tumor is preferred. The kit according to this aspect is suitably used for the test methods according to the above-described first to third aspects.

[0169] The reagent for measuring free fatty acids is a reagent used 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 free fatty acids such as 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 in group A1). For example, the reagent for measuring free fatty acids may be an antibody or an aptamer that specifically binds to a specific free fatty acid.

[0170] When measuring free fatty acids in a sample using analytical instruments such as gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, and gas chromatography, examples of the reagent for measuring the concentration of free fatty acids include standard samples of any of the above free fatty acids. The standard sample is used for creating a calibration curve in the measurement of free fatty acids by the above analytical instruments. The concentration of free fatty acids in the sample can be calculated based on the calibration curve created with the standard sample. The standard sample of free fatty acids may be a purified product of the above free fatty acids. The purity of the specific free fatty acid in the standard sample of the specific free fatty acid is, for example, 96% by mass or more, 97% by mass or more, 98% by mass or more, or 99% by mass or more.

[0171] The test kit according to this embodiment only needs to contain a reagent for measuring any one or more of the free fatty acids in the above A1 group, and may contain a reagent for measuring 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, fourteen or more of the free fatty acids in the above A1 group. The fatty acid that is the measurement target of the reagent that the test kit according to this embodiment may contain can be arbitrarily selected from the free fatty acids in the A1 group. Alternatively, the test kit according to this embodiment may contain a reagent for measuring all 15 kinds of the free fatty acids in the above A1 group.

[0172] When the kit according to this embodiment is a gynecological cancer test kit, it preferably contains 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, and docosapentaenoic acid. The kit according to this embodiment can contain a reagent 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, twelve or more, fourteen or more, or 14 of the above free fatty acids.

[0173] When the kit according to this embodiment is an ovarian cancer test kit, it preferably contains 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 according to this embodiment can contain a reagent 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 above free fatty acids.

[0174] When the kit according to this embodiment is also intended to detect early gynecological cancers (e.g., Stage I, Stage II), it 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 lignoceric acid. The kit according to this embodiment can include a reagent for measuring 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, or 8 of the above free fatty acids.

[0175] When the kit according to this embodiment is a test kit for endometrial cancer or cervical cancer, it preferably includes 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 according to this embodiment can include a reagent 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 above free fatty acids.

[0176] When the kit according to this embodiment is a test kit for endometrial cancer, it preferably includes 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 according to this embodiment can include a reagent for measuring 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, or 8 of the above free fatty acids.

[0177] When the kit according to this embodiment is a cervical cancer test kit, it preferably includes 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 according to this embodiment can include a reagent 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 above free fatty acids.

[0178] When the kit according to this embodiment is a test kit used for the evaluation in (b) above, it 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, 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 according to this embodiment can include a reagent 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, twelve or more, thirteen or more, or fourteen of the above free fatty acids.

[0179] When the kit according to this embodiment is an evaluation test kit for the possibility of a poor prognosis in a subject with ovarian cancer, it 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, adrenic acid, nervonic acid, and lignoceric acid. The kit according to this embodiment can include a reagent 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 above free fatty acids.

[0180] When the kit according to this embodiment is an evaluation test kit for the possibility of poor prognosis in a subject having endometrial cancer or cervical cancer, it preferably includes 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 according to this embodiment can include a reagent 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 above free fatty acids.

[0181] When the kit according to this embodiment is an evaluation test kit for the possibility of poor prognosis in a subject having endometrial cancer, it preferably includes 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 according to this embodiment can include a reagent for measuring three or more, four or more, five or more, six or more, seven or more, or eight of the above free fatty acids.

[0182] When the kit according to this embodiment is an evaluation test kit for the possibility of poor prognosis in a subject having cervical cancer, it preferably includes 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 according to this embodiment can include a reagent 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 above free fatty acids.

[0183] When the kit according to this embodiment is an inspection kit used for the evaluation in the above (c), it 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, 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 according to this embodiment can include a reagent 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, twelve or more, thirteen or more, or fourteen of the above free fatty acids.

[0184] When the kit according to this embodiment is an evaluation inspection kit for evaluating whether the ovarian cancer possessed by the subject is a malignant tumor or a borderline malignant tumor, it 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, adrenic acid, nervonic acid, and lignoceric acid. The kit according to this embodiment can include a reagent 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 above free fatty acids.

[0185] When the kit according to this embodiment is an evaluation inspection kit for evaluating whether the endometrial cancer or cervical cancer possessed by the subject is a malignant tumor or a borderline malignant tumor, it preferably includes 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 according to this embodiment can include a reagent 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 above free fatty acids.

[0186] When the kit according to this embodiment is an evaluation test kit for evaluating whether the uterine body cancer possessed by the subject is a malignant tumor or a borderline malignant tumor, it preferably includes 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 according to this embodiment can include a reagent for measuring three or more, four or more, five or more, six or more, seven or more, or eight of the above free fatty acids.

[0187] When the kit according to this embodiment is an evaluation test kit for evaluating whether the cervical cancer possessed by the subject is a malignant tumor or a borderline malignant tumor, it preferably includes 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 according to this embodiment can include a reagent 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 above free fatty acids.

[0188] When the kit according to this embodiment is a test kit used for the evaluation in (d) above, it preferably includes a reagent for measuring one or more free fatty acids selected from the group consisting of dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid. The kit according to this embodiment can include a reagent for measuring two or more, three or more, or four of the above free fatty acids.

[0189] When measuring free fatty acids in a sample using analytical instruments such as gas chromatography - mass spectrometry, liquid chromatography - mass spectrometry, and gas chromatography, the kit according to this embodiment may contain an internal standard substance. The internal standard substance is added to and used with a sample derived from the subject. The concentration of free fatty acids in the sample can be calculated from the ratio of the added amount of the internal standard substance added to the sample derived from the subject to the measured value of the free fatty acids relative to the measured value of the internal standard substance in the sample. As the internal standard substance, a substance not contained in the sample derived from the subject can be used. For example, a free fatty acid 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 similar to that of the free fatty acids to be measured with respect to the solvent used when extracting free fatty acids from the sample derived from the subject. Examples of such internal standard substances include margaric acid.

[0190] By using the kit according to this embodiment, it becomes possible to simply measure the concentration of the above - mentioned free fatty acids in a sample derived from the subject. As a result, it is possible to simply obtain the evaluation results of the inspection methods according to the first to third aspects.

[0191] [Medicine] The medicine according to this aspect is a medicine for treating or preventing gynecological cancer, which is administered to a subject evaluated as having gynecological cancer by the inspection method according to the first aspect.

[0192] Examples of the gynecological cancer include the cancers described above in the first aspect. The gynecological cancer is preferably a gynecological malignant tumor, more preferably a malignant tumor selected from the group consisting of ovarian malignant tumor, uterine body malignant tumor, and cervical malignant tumor, and even more preferably ovarian malignant tumor.

[0193] The medicament according to this aspect is not particularly limited as long as it is effective against gynecological cancer. Examples of medicaments for gynecological cancer include known anticancer agents used in the treatment of gynecological cancer. Such anticancer agents include, for example, carboplatin, cisplatin, paclitaxel, docetaxel, irinotecan, doxil, topotecan, gemcitabine, etoposide, trabectedin, abraxane, tamoxifen, ifosfamide, 5-FU, doxorubicin, and the like. Alternatively, the medicament according to this embodiment may be an antibody medicament, and examples thereof include known antibody medicaments. Examples of antibodies that can be contained in the antibody medicament include anti-VEGF antibodies such as bevacizumab.

[0194] The medicament according to this aspect may contain various substances generally used for formulation, such as excipients, stabilizers, other pharmaceutically acceptable components, or other pharmaceutically active ingredients.

[0195] The medicament according to this aspect may be administered orally or parenterally. Depending on the administration route, the medicament according to this embodiment can take various dosage forms, such as solid preparations, liquid preparations, and the like. For example, it can be an oral preparation such as an oral solid preparation or an oral liquid preparation, or a parenteral preparation such as an injection or an infusion. Examples of carriers that can be used for parenteral preparations include aqueous carriers such as physiological saline and isotonic solutions containing glucose or D-sorbitol.

[0196] The dosage of the medicament according to this aspect varies depending on the age, weight, disease to be treated, symptoms, etc. of the subject. For example, as a single dose, it can be about 0.1 μg to 1 mg / kg per body weight. The dosing interval is not particularly limited, and for example, it can be once a day to once every few months.

[0197] [Other aspects] In one aspect, the present disclosure provides a biomarker for evaluating the presence or absence of one or more selected from the group consisting of gynecological cancers and their precancerous lesions, 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 aspect, the present disclosure provides a biomarker for predicting the prognosis of a gynecological cancer patient, 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 aspect, the present disclosure provides a biomarker for classifying gynecological malignancies and gynecological borderline malignancies, 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 aspect, the present disclosure provides a biomarker for classifying gynecological cancers and their precancerous lesions, selected from the group consisting of dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid.

Example

[0198] Hereinafter, the present invention will be described by way of examples, but the present invention is not limited to the following examples. In the examples, ovarian cancer, endometrial cancer, and cervical cancer mean ovarian malignant tumor, endometrial malignant tumor, and cervical malignant tumor, respectively.

[0199] [Experimental Example 1] (Expression analysis of fatty acid metabolism enzymes) The expression levels of fatty acid metabolism enzymes in normal ovarian tissue and ovarian cancer tissue were analyzed by RT-PCR.

[0200] Nine cases of normal ovarian tissue were collected during surgery for gynecological malignancies for therapeutic purposes (such as prevention of metastasis). Thirty-two cases of ovarian cancer tissue were collected during surgery for ovarian malignancies. Each normal ovarian tissue and ovarian cancer tissue was pulverized with a crusher, and then total RNA was extracted using the RNeasy Kit (QIAGEN).

[0201] cDNA synthesis was performed using ReverTra Ace qPCR RT Master Mix with gDNA Remover (manufactured by TOYOBO). Specifically, first, 1 μg equivalent of total RNA and RNase free distilled water were mixed to a volume of 12 μL and incubated at 65 °C for 5 minutes to denature the RNA. Subsequently, on ice, 4 μL of 4×DN Master Mix and gDNA Remover were added and reacted at 37 °C for 5 minutes to remove genomic DNA. Subsequently, 4 μL of 5×RT Master Mix II was further mixed, and a reverse transcription reaction (37 °C for 15 minutes, 50 °C for 5 minutes, 98 °C for 5 minutes) was performed to synthesize cDNA.

[0202] Using the cDNA prepared above as a template, real-time PCR was performed using THUNDERBIRD Probe qPCR Mix (manufactured by TOYOBO). Real-time PCR was carried out according to the 2 -ΔCt protocol. That is, the difference in Ct values between the target gene and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was taken as the ΔCt value, and the relative quantification value was calculated by 2 -ΔCt . Each measurement was performed in triplicate. The target genes were human acetyl-CoA carboxylase 1 (ACACA), fatty acid synthase (FASN), stearoyl-CoA desaturase 1 (SCD1), fatty acid desaturase 1 (FADS1), FADS2, elongation of very long chain fatty acid 1 (ELOVL1), ELOVL2, ELOVL3, ELOVL4, ELOVL5, and ELOVL6.

[0203] The results are shown in Figures 1A to 1D. Figure 1A is a diagram showing the expression levels of free fatty acid-metabolizing enzymes in normal ovarian tissue and ovarian cancer tissue by a heat map. The heat map was created using MetaboAnalyst 5.0 software.

[0204] Figures 1B to 1D are graphs showing the expression levels of each fatty acid-metabolizing 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 a t-test, and p < 0.05 was considered to be significant. "*", "**", "***", "****" indicate p < 0.05, p < 0.01, p < 0.001, and p < 0.0001, respectively.

[0205] From the results shown in Figure 1B, it was confirmed that the expression levels of FASN and SCD1 were higher in ovarian cancer tissue than in normal ovarian tissue. From the results shown in Figure 1C, it was confirmed that the expression levels of ELOVL1, ELOVL2, ELOVL3, ELOVL4, ELOVL5, ELOVL6, and FADS1 were lower in ovarian cancer tissue than in normal ovarian tissue. From the results shown in Figure 1D, it was confirmed that the expression levels of ACACA and FADS2 were comparable between normal ovarian tissue and ovarian cancer tissue. From the above results, it was confirmed that the expression levels of fatty acid-metabolizing enzymes differed between normal ovarian tissue and ovarian cancer tissue.

[0206] [Experimental Example 2] (Free Fatty Acid Concentration in Serum of Ovarian Cancer Patients) The amount of free fatty acids in the sera of healthy women and ovarian cancer patients was analyzed by gas chromatography-mass spectrometry (GC-MS).

[0207] Blood samples were collected 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) to obtain serum samples.

[0208] First, a calibration curve was created using standard samples for measuring free fatty acids in serum. As the 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), sapienic acid (manufactured by Cayman Chemical, #9001845), vaccenic acid (manufactured by Cayman Chemical, #20023), and margaric acid (manufactured by Cayman Chemical, Nacalai Tesque) as the internal standard substance were used.

[0209] First, 20 μL of healthy subject serum and ovarian cancer patient serum were diluted with 300 μL of phosphate-buffered saline (PBS) containing 100 ng of margaric acid as the internal standard substance. Subsequently, the entire volume of the sample was applied to a diatomaceous earth column (product name "SLE+400", Biotage) and eluted with 1.8 mL of dichloromethane. Subsequently, the eluate was dried under a nitrogen stream, and the obtained dried product was dissolved in 5 μL of pyridine. 30 μL of BSTFA:TMCS (99:1, ThermoFisher TS-38831) was added thereto and mixed to trimethylsilylate all OH groups.

[0210] Subsequently, 2 μL of the derivatized sample was injected into the GC-MS for analysis and interpretation. The quantitative values were determined by comparing the peak area ratios of the internal standard substance and each free fatty acid with the calibration curve prepared in advance.

[0211] The analysis conditions of GC-MS were as follows. As the capillary column, RTx-5MS (length 30 m, inner diameter 0.25 mm, film thickness 0.25 μm, manufactured by Restek) was used. The oven temperature was held at 150 °C for 1 minute, heated to 250 °C at a rate of 20 °C / min, then heated to 280 °C at a rate of 5 °C / min, held at 280 °C for 3 minutes, heated to 330 °C at a rate of 20 °C / min, and held at 330 °C for 1 minute. He was used as the carrier gas. The linear velocity was set at 42.0 cm / sec. The ion source temperature, interface temperature, and vaporization chamber temperature were set at 200 °C, 280 °C, and 250 °C, respectively.

[0212] The free fatty acids to be measured were 18 types, namely 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, nervonic acid, and lignoceric acid.

[0213] The measurement results are shown in FIGS. 2A to 2D. FIG. 2A is a diagram showing the average values of the amounts of the various free fatty acids in the sera of healthy subjects and ovarian cancer patients as a heatmap. The heatmap was created using MetaboAnalyst 5.0 software.

[0214] FIGS. 2B to 2C are graphs showing the individual values of the various free fatty acids in the sera of healthy subjects and ovarian cancer patients. The comparison between the two groups was performed by a t-test, and p < 0.05 was considered to indicate a significant difference. "*", "**", "***", and "****" indicate p < 0.05, p < 0.01, p < 0.001, and p < 0.0001, respectively.

[0215] Figure 2D is a volcano plot showing the measurement results of each free fatty acid. In Figure 2D, "Up" represents free fatty acids with significantly higher concentrations in ovarian cancer patient serum than in healthy subject serum. "Down" represents free fatty acids with significantly lower concentrations in ovarian cancer patient serum than in healthy subject serum. "non-sig" represents free fatty acids for which no significant difference in concentration was confirmed between healthy subject serum and ovarian cancer patient serum. The horizontal axis represents the ratio of the average value of the concentration of each free fatty acid in ovarian cancer patient serum to the average value of the concentration of each free fatty acid in healthy subject serum. The vertical axis represents the P value when there is no difference in the concentration of each free fatty acid between healthy subject serum and ovarian cancer patient serum.

[0216] From the results shown in Figures 2A - C, it was confirmed that the concentrations of palmitoleic acid, palmitic acid, linoleic acid, α-linolenic acid, and oleic acid were higher in ovarian cancer patient serum than in healthy subject serum. Conversely, it was confirmed that the concentrations of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, and lignoceric acid were lower in ovarian cancer patient serum than in healthy subject serum.

[0217] The free fatty acids with significant differences between healthy subject serum and ovarian cancer patient serum were summarized in Table 1. In Table 1, the description "high" in the concentration difference column means that the concentration of that free fatty acid was significantly higher in ovarian cancer patient serum than in healthy subject serum. In Table 1, the description "low" in the concentration difference column means that the concentration of that free fatty acid was significantly lower in ovarian cancer patient serum than in healthy subject serum.

[0218] [Table 1]

[0219] [Experimental Example 3] (Correlation Analysis between Expression Levels of Fatty Acid Metabolizing Enzymes and Concentrations of Free Fatty Acids) The correlation between the expression levels of fatty acid metabolizing enzymes in tissues and the concentrations of free fatty acids in serum was analyzed.

[0220] Figures 3A - 3B show the correlation between the changes in the expression of fatty acid metabolic enzymes in the tissues shown in Figures 1B - D and the changes in the free fatty acid composition in the sera shown in Figures 2B - C. "HIGH" refers to the metabolic enzymes whose expression was significantly higher in ovarian cancer tissues than in normal ovarian tissues in Figures 1B - D. "LOW" refers to the metabolic enzymes whose expression was significantly lower in ovarian cancer tissues than in normal ovarian tissues in Figures 1B - D. "not significant" refers to the metabolic enzymes for which no significant difference in expression was confirmed between normal ovarian tissues and ovarian cancer tissues in Figures 1B - D.

[0221] Ovarian cancer tissues and serum samples were collected in pairs from 20 ovarian cancer patients at the same time. The correlation between the expression level of SCD1 in ovarian cancer tissues and the ratio of free fatty acids that are substrates of SCD1 and the amount ratio of free fatty acids generated by the action of SCD1 in this paired sample was analyzed.

[0222] SCD1 acts on palmitic acid as a substrate to produce palmitoleic acid. Also, SCD1 acts on stearic acid to produce oleic acid.

[0223] Figure 3C is a graph showing the correlation between the SCD1 gene expression level in ovarian cancer tissues and the ratio of free fatty acids corresponding to SCD1 activity in the sera of ovarian cancer patients (generated free fatty acids / substrate free fatty acids ratio (palmitoleic acid / palmitic acid ratio)). Figure 3D is a graph showing the correlation between the SCD1 gene expression level in ovarian cancer tissues and the ratio of free fatty acids corresponding to SCD1 activity in the sera of ovarian cancer patients (generated free fatty acids / substrate free fatty acids ratio (oleic acid / stearic acid ratio)). The correlation analysis was performed by Pearson's correlation test using GraphPad Software version 9.0, and the r - value and p - value were calculated. The results shown in Figures 3A - 3D indicate that the changes in the expression of fatty acid metabolic enzymes in tissues and the changes in the concentration of free fatty acids in sera are linked.

[0224] [Experimental Example 4] (ROC Analysis for Detecting Ovarian Cancer Patients by Free Fatty Acid Concentration) In Experimental Example 2, Receiver operating characteristic analysis (ROC analysis) was performed using 11 free fatty acids for which a significant difference was confirmed in the serum concentrations between healthy women and ovarian cancer patients as indices.

[0225] ROC analysis was performed for healthy subjects and ovarian cancer patients using the concentrations of each free fatty acid. Figure 4A shows the results of ROC analysis using the concentrations of palmitoleic acid, palmitic acid, linoleic acid, α-linolenic acid, and oleic acid. Figure 4B shows the results of ROC analysis using the concentrations of vaccenic acid, arachidic acid, docosahexaenoic acid, adrenic acid, nervonic acid, and lignoceric acid. In Figures 4A to 4B, AUC and CI mean Area Under Curve and Confidence Interval, respectively.

[0226] Among these 11 types, it was revealed that 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, nervonic acid, and lignoceric acid, it was possible to distinguish whether the subject of the test was a healthy subject or an ovarian cancer patient.

[0227] [Experimental Example 5] (ROC Analysis for Detection of Ovarian Cancer Patients by FFA Score) For the 10 free fatty acids for which the results of the ROC analysis in Experimental Example 4 were good, a threshold was set so that the positive likelihood ratio in the ROC analysis in Experimental Example 4 was maximized. The threshold for each free fatty acid was 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 nervonic acid.

[0228] For the subjects analyzed in Experimental Example 2, based on the thresholds of each of the 10 free fatty acids set above, it was determined whether the serum concentration of each free fatty acid was an abnormal value. The results for healthy subjects are shown in Fig. 5A. The results for ovarian cancer patients are shown in Fig. 5B. In Figs. 5A to 5B, "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 nervonic acid. For the free fatty acids of "1" to "4", when the measured concentration was higher than the threshold, the concentration was determined to be an abnormal value and indicated by a black circle, and when the measured concentration was lower than the threshold, the concentration was determined to be a normal value and indicated by a white circle. For the free fatty acids of "5" to "10", when the measured concentration was lower than the threshold, the concentration was determined to be an abnormal value and indicated by a black circle, and when the measured concentration was higher than the threshold, the concentration was determined to be a normal value and indicated by a white circle.

[0229] From the determination results shown in Figs. 5A to 5B, the number of black circles for each subject was calculated as the FFA score of each subject. Using this FFA score, ROC analysis was performed for healthy female subjects and ovarian cancer patients. The results of the ROC analysis are shown in Fig. 5C. As a result of the ROC analysis, the AUC value was 0.9963. The CI value was 0.9880 - 1.000. The p value was P < 0.0001. From this result, it was confirmed that ovarian cancer patients can be detected with high sensitivity by using this FFA score.

[0230] [Experimental Example 6] (Free Fatty Acid Concentrations in Serum of Early Ovarian Cancer Patients) The serum concentrations of 10 free fatty acids, for which the results of ROC analysis were good in Experimental Example 4, were compared among 27 healthy female subjects, 29 ovarian cancer patients at Stage I or Stage II, and 11 ovarian cancer patients at Stage III or Stage IV.

[0231] The results of the comparison among the three groups are shown in FIGS. 6A and 6B. In the figures, "HD" means healthy female subjects. The comparison among the three groups was performed by one-way ANOVA and Bonferroni's multiple comparisons tests, and a p < 0.05 was considered to indicate a significant difference. "*", "**", "***", and "****" indicate p < 0.05, p < 0.01, p < 0.001, and p < 0.0001, respectively.

[0232] From the results shown in FIG. 6A, it was confirmed that the concentrations of four types of free fatty acids, palmitoleic acid, linoleic acid, α-linolenic acid, and oleic acid, were significantly higher in the sera of ovarian cancer patients at Stage I or Stage II than in healthy subject sera. From the results shown in FIG. 6B, it was confirmed that the concentrations of four types of free fatty acids, vaccenic acid, arachidic acid, docosahexaenoic acid, and lignoceric acid, were significantly lower in the sera of ovarian cancer patients at Stage I or Stage II than in healthy subject sera. It was confirmed that the serum concentrations of the above eight types of free fatty acids can be an index for distinguishing between healthy subjects and ovarian cancer patients at Stage I or Stage II. In addition, in ovarian cancer patients at Stage III or Stage IV, no significant difference was confirmed in the serum concentrations of α-linolenic acid, docosahexaenoic acid, and lignoceric acid compared with healthy subjects.

[0233] [Experimental Example 7] (ROC Analysis for Detection of Early Ovarian Cancer Patients by Free Fatty Acid Concentrations) In Experimental Example 6, Receiver operating characteristic analysis (ROC analysis) was performed using the serum concentrations of eight types of free fatty acids for which significant differences were confirmed in the serum concentrations between healthy female subjects and ovarian cancer patients at Stage I or Stage II as indices.

[0234] Using the concentrations of each free fatty acid, ROC analysis was performed in healthy subjects and ovarian cancer patients at Stage I or Stage II. Figure 7A shows the results of ROC analysis using the concentrations of palmitoleic acid, linoleic acid, α-linolenic acid, and oleic acid. Figure 7B shows the results of ROC analysis using the concentrations of vaccenic acid, arachidic acid, docosahexaenoic acid, and lignoceric acid. In Figures 7A - B, AUC and CI respectively mean Area Under Curve and Confidence Interval.

[0235] Based on the concentrations of these 8 free fatty acids, it was revealed that it was possible to distinguish whether the subject of the test was a healthy subject or an ovarian cancer patient at Stage I or Stage II.

[0236] [Experimental Example 8] (ROC Analysis by FFA Score: Detection of Early Ovarian Cancer Patients) For the 8 free fatty acids subjected to ROC analysis in Experimental Example 7, the threshold was set so that the positive likelihood ratio in the ROC analysis in Experimental Example 7 was maximized. The threshold for each free fatty acid was 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 lignoceric acid.

[0237] For the subjects analyzed in Experimental Example 6, based on the thresholds of the eight free fatty acids set above, it was determined whether the serum concentration of each free fatty acid was an abnormal value. The results for healthy subjects are shown in FIG. 8A. The results for ovarian cancer patients at Stage I or Stage II are shown in FIG. 8B. In FIGS. 8A to 8B, "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. For the free fatty acids "1" to "4", when the measured concentration was higher than the threshold, the concentration was determined to be an abnormal value and indicated by a black circle, and when the measured concentration was lower than the threshold, the concentration was determined to be a normal value and indicated by a white circle. For the free fatty acids "5" to "8", when the measured concentration was lower than the threshold, the concentration was determined to be an abnormal value and indicated by a black circle, and when the measured concentration was higher than the threshold, the concentration was determined to be a normal value and indicated by a white circle.

[0238] From the determination results shown in FIGS. 8A to 8B, the number of black circles for each subject was calculated as the FFA score of each subject. Using this FFA score, ROC analysis was performed for healthy female subjects and ovarian cancer patients at Stage I and Stage II. The results are shown in FIG. 8C. As a result of the ROC analysis, 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, ovarian cancer patients at Stage I or Stage II can be detected with high sensitivity.

[0239] The threshold of the FFA score was set so that the positive likelihood ratio of the ROC analysis in FIG. 8C was maximized. The threshold of the FFA score was 1.5.

[0240] Using the above-mentioned FFA score threshold, the detection sensitivity by the FFA score for ovarian cancer in Stage I or II was compared with the detection sensitivity by the blood concentration of CA125. Figure 8D shows the FFA score (left figure) and the blood concentration of CA125 (right figure) in patients with Stage I or Stage II ovarian cancer. In Figure 8D, "×" indicates clear cell type ovarian cancer, and "●" indicates ovarian cancer other than clear cell carcinoma type.

[0241] From the analysis results of Figure 8D, when the threshold of the FFA score was set at 1.5, the detection sensitivity for patients with Stage I or Stage II ovarian cancer was 100%. When the threshold of the blood concentration of CA125 was set at 35 U / mL used in clinical practice, the detection sensitivity for patients with Stage I or Stage II ovarian cancer was 76%. From these results, it was confirmed that by using the FFA score calculated from 8 free fatty acid values, patients with Stage I or Stage II ovarian cancer can be detected with higher sensitivity than by examining using the blood concentration of CA125. Furthermore, it was confirmed that the FFA score can also detect clear cell type ovarian cancer, which is a tissue type difficult to detect by CA125, with high sensitivity.

[0242] [Experimental Example 9] (Prognosis prediction of ovarian cancer patients)

[0243] [[ID=I4]]For the 40 ovarian cancer patients 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 40 ovarian cancer patients, patients with an FFA score higher than the median value were classified into the "FFA score high" group, and patients with an FFA score below the median value were classified into the "FFA score low" group. Note that the serum samples used for calculating the FFA score in Experimental Example 5 were obtained by blood sampling before the start of treatment. The prognosis of the patients in the FFA score high group and the patients in the FFA score low group was followed up respectively, and the transition of the progression-free survival rate in each patient group was examined. The results are shown in Figure 9. In Figure 9, 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 for the significance test.

[0244] It was confirmed that the treatment prognosis of ovarian cancer patients can be predicted using the FFA score as an indicator.

[0245] [Experimental Example 10] (ROC analysis using the FFA score for the detection of borderline malignant ovarian tumors) Blood was collected from 7 patients with borderline malignant ovarian tumors to obtain serum samples. Subsequently, the serum concentrations of each free fatty acid were measured by GC-MS in the same manner as in Experimental Example 2. The free fatty acids to be measured were the 10 free fatty acids analyzed in Experimental Example 5. Subsequently, the FFA score of each patient with borderline malignant ovarian tumor was calculated in the same manner as in Experimental Example 5.

[0246] The threshold values of each free fatty acid used for calculating 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 nervonic acid was 0.0891 μEQ / L.

[0247] Figure 10A shows the results of determining whether the serum concentration of each free fatty acid is an abnormal value for patients with borderline malignant tumors based on the above threshold values.

[0248] Using the FFA score of patients with borderline malignant ovarian tumors calculated from the results in Figure 10A and the FFA score of ovarian cancer patients (patients with malignant ovarian tumors) calculated from the results in Figure 5B, ROC analysis was performed. The results are shown in Figure 10B. As a result of the 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 this result, it was confirmed that by using the FFA score, patients with borderline malignant ovarian tumors and ovarian cancer patients (patients with malignant ovarian tumors) can be distinguished with high sensitivity.

[0249] [Experimental Example 11] (Analysis of endometrial cancer and cervical cancer) Blood samples were collected from 27 healthy adult women, 66 endometrial cancer patients, and 45 cervical cancer patients to obtain serum samples. Subsequently, the serum concentrations of each free fatty acid were measured by GC-MS in the same manner as in Experimental Example 2.

[0250] The measurement results are shown in FIGS. 11A - B. The comparison among the three groups was performed by one-way ANOVA and Bonferroni’s multiple comparisons tests, and p < 0.05 was considered to indicate a significant difference. “**” and “****” indicate p < 0.01 and p < 0.0001, respectively.

[0251] The concentrations of palmitoleic acid, palmitic acid, linoleic acid, α-linolenic acid, and oleic acid were confirmed to be higher in the sera of endometrial cancer and cervical cancer patients than in the sera of healthy subjects. The concentrations of vaccenic acid, arachidic acid, and stearic acid were confirmed to be lower in the sera of endometrial cancer patients and cervical cancer patients than in the sera of healthy subjects. From these results, it became clear that the serum concentrations of these free fatty acids can be used as an index to distinguish whether the subject of the test is a healthy person, an endometrial cancer patient, or a cervical cancer patient.

[0252] [Experimental Example 12] (Free fatty acid concentrations in the sera of early-stage cervical cancer patients) Blood samples were 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. Subsequently, the serum concentrations of each free fatty acid were measured by GC-MS in the same manner as in Experimental Example 2.

[0253] The measurement results are shown in FIGS. 12A - C. The comparison among the three groups was performed by one-way ANOVA and Bonferroni’s multiple comparisons tests, and a p < 0.05 was considered to indicate a significant difference. "*", "**", "***", and "****" indicate p < 0.05, p < 0.01, p < 0.001, and p < 0.0001, respectively.

[0254] The concentrations of palmitoleic acid, linoleic acid, α-linolenic acid, and oleic acid were confirmed to be significantly higher in the sera of Stage I cervical cancer patients or Stage II or III cervical cancer patients than in the sera of healthy subjects. On the other hand, the concentrations of vaccenic acid, arachidic acid, dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid were confirmed to be significantly lower in the sera of Stage I cervical cancer patients or Stage II or III cervical cancer patients than in the sera of healthy subjects. It was confirmed that the serum concentrations of these free fatty acids can be used as an index to distinguish between healthy subjects and Stage I, Stage II, or Stage III cervical cancer patients.

[0255] Furthermore, the threshold values of the concentrations of each free fatty acid were set to the values shown in Table 2, and ROC analysis was performed using the concentrations of each free fatty acid in healthy subjects and Stage I cervical cancer patients. The results are shown in FIGS. 12A - C. The AUC of each free fatty acid was 0.7 or more. It was clarified that the serum concentrations of these free fatty acids can be used as an index to distinguish whether the test subject is a healthy subject or a Stage I cervical cancer patient.

[0256]

Table 2

[0257] [Experimental Example 13] (Analysis of the ratio of free fatty acid concentrations in the sera of early-stage cervical cancer patients) Using the concentration ratio of 10 free fatty acids as an index, it was verified whether it was possible to distinguish whether the subject of the examination was a healthy person or a cervical cancer patient.

[0258] For the serum samples of Experimental Example 12, the ratio of docosapentaenoic acid to oleic acid (docosapentaenoic acid / oleic acid) was calculated. The calculation results are shown in Fig. 13A. The comparison among the three groups was performed by one-way ANOVA and Bonferroni’s multiple comparisons tests, 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.

[0259] The ratio (docosapentaenoic acid / oleic acid) was confirmed to be significantly lower in the sera of Stage I cervical cancer patients or Stage II or III cervical cancer patients than in the sera of healthy subjects. The ratio (docosapentaenoic acid / oleic acid) was confirmed to be able to serve as an index for distinguishing between healthy subjects and Stage I, Stage II, or Stage III cervical cancer patients.

[0260] Furthermore, the threshold value 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) to distinguish between healthy subjects and Stage I cervical cancer patients. The results are shown in Fig. 13A. The ROC analysis results were AUC = 0.981, P<0.0001. That is, it was confirmed that by using the ratio (docosapentaenoic acid / oleic acid) as an index rather than using either docosapentaenoic acid or oleic acid as an index, it was possible to distinguish with higher accuracy whether the subject of the examination was a healthy person or a Stage I cervical cancer patient.

[0261] Next, for the 29 Stage I cervical cancer patients who were the subjects of the examination in Experimental Example 12, the detection sensitivity by the ratio (docosapentaenoic acid / oleic acid) was calculated and compared with the detection cancer rate by existing diagnostic markers. The results are shown in Fig. 13B.

[0262] When the threshold of the ratio (docosahexaenoic acid / oleic acid) was set at 0.004433, the detection sensitivity for Stage I cervical cancer patients was 93.1%. When the threshold of the blood concentration of SCC was set at 1.5 ng / mL, which is used in actual clinical practice, the detection sensitivity for Stage I cervical cancer patients was 68.2%. When the threshold of the blood concentration of CEA was set at 5.0 ng / mL, which is used in actual clinical practice, the detection sensitivity for Stage I cervical cancer patients was 4.2%.

[0263] From these results, it was confirmed that by using the ratio (docosahexaenoic acid / oleic acid), Stage I cervical cancer patients can be detected with higher sensitivity than by examining using the blood concentration of SCC or CEA, which are existing diagnostic markers.

[0264] [Experimental Example 14] (Analysis of Patients with Cervical Precancerous Lesions) Blood samples were collected from 27 healthy adult women and 17 patients with cervical precancerous lesions (cervical dysplasia: CIN) to obtain serum samples. Subsequently, the serum concentrations of each free fatty acid were measured by GC-MS in the same manner as in Experimental Example 2.

[0265] The measurement results are shown in FIGS. 14A - B. The comparison among the three groups was performed by one-way ANOVA and Bonferroni’s multiple comparisons tests, and p < 0.05 was considered to indicate a significant difference. “*”, “**”, “***”, and “****” indicate p < 0.05, p < 0.01, p < 0.001, and p < 0.0001, respectively.

[0266] It was confirmed that the concentrations of palmitoleic acid, linoleic acid, α-linolenic acid, oleic acid, and arachidonic acid were significantly higher in the serum of CIN patients than in the serum of healthy subjects. On the other hand, it was confirmed that the concentration of arachidic acid was significantly lower in the serum of CIN patients than in the serum of healthy subjects. It was confirmed that the serum concentrations of these free fatty acids can be an index for distinguishing between healthy subjects and CIN patients.

[0267] Furthermore, the threshold values of the concentrations of the respective free fatty acids were set to the values shown in Table 3, and ROC analysis was performed using the concentrations of the respective free fatty acids in healthy subjects and CIN patients. The results are shown in FIGS. 14A to 14B. The AUC of each free fatty acid was 0.7 or more. It was clarified that the serum concentrations of these free fatty acids can be used as an index to distinguish whether the subject of the test is a healthy subject or a CIN patient.

[0268]

Table 3

[0269] [Experimental Example 15] (Analysis of CIN patients and cervical cancer patients) Regarding the sera 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 subject of the test was a CIN patient or a cervical cancer patient.

[0270] 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 tests, and p < 0.05 was considered to indicate a significant difference. "***" and "****" indicate p < 0.001 and p < 0.0001, respectively.

[0271] It was confirmed that the concentrations of dihomo-γ-linolenic acid, docosahexaenoic acid, adrenic acid, and docosapentaenoic acid were significantly lower in the sera of cervical cancer patients than in the sera of CIN patients. It was confirmed that the serum concentrations of these free fatty acids can be used as an index to distinguish between CIN patients and cervical cancer patients.

[0272] Furthermore, the threshold values of the concentrations of the respective free fatty acids were set to the values shown in Table 4, and ROC analysis was performed using the concentrations of the respective free fatty acids in CIN patients and cervical cancer patients. The results are shown in Fig. 15. The AUC of each free fatty acid was 0.7 or more. It was revealed that the serum concentrations of these free fatty acids can be used as an index to distinguish whether the test subject is a CIN patient or a cervical cancer patient.

[0273]

Table 4

Industrial Applicability

[0274] According to the present invention, it is possible to provide a simple and highly accurate test method for detecting one or more selected from the group consisting of gynecological cancers and their precancerous lesions, predicting the prognosis of gynecological cancers, and classifying gynecological malignancies and gynecological borderline malignancies, a medicament for treating a subject determined to have a gynecological cancer by the test method, and a kit. The present invention is useful for early diagnosis of gynecological cancers or their precancerous lesions.

Claims

1. 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, dihomo-γ-linolenic acid, and docosapentaenoic acid in a sample derived from a subject; A step (B1) of identifying a sample derived from a subject who is highly likely to have one or more selected from the group consisting of gynecological cancers and their precancerous lesions, based on the concentration of the free fatty acids in the sample obtained in the step (A1); In the step (B1), a method for examining a sample, wherein when at least one of the following (i1), (ii1), and (iii1) is satisfied, the sample is identified as being derived from a subject who is highly likely to have one or more selected from the group consisting of gynecological cancers and their precancerous lesions; (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, and palmitic acid in the sample is high as compared with 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, dihomo-γ-linolenic acid, and docosapentaenoic acid in the sample is low as compared with a predetermined threshold value; (iii1) The concentration ratio of docosapentaenoic acid to oleic acid is low as compared with a predetermined threshold value.

2. 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 a gynecological cancer; A step (B2) of identifying a sample derived from a subject who is highly likely to have a poor prognosis, based on the concentration of the free fatty acids in the sample obtained in the step (A2); In the step (B2), a method for examining a sample, wherein when at least one of the following (i2) and (ii2) is satisfied, the sample is identified as being derived from a subject who is highly likely to have 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 high as compared with 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 low as compared with a predetermined threshold value.

3. 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 a gynecological cancer; A step (B3) of evaluating whether the sample is derived from a subject highly likely to have a gynecological malignant tumor or a subject highly likely to have a borderline gynecological malignant tumor based on the concentration of the free fatty acid in the sample obtained in the step (A3), the method comprising: A method for examining a sample, wherein in the step (B3), when at least one of the following (i3) and (ii3) is satisfied, the sample is identified as being derived from a subject highly likely to have a gynecological 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 high as compared with 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 low as compared with a predetermined threshold value.

4. Based on the concentration of the free fatty acids in the sample obtained in the step (A1), further comprising a step (C1) of evaluating whether the sample identified as being derived from a subject highly likely to have one or more selected from the group consisting of gynecological cancers and their precancerous lesions is derived from a subject highly likely to have a gynecological cancer or a subject highly likely to have a precancerous lesion of a gynecological cancer. The method for examining a sample according to claim 1, wherein in the step (C1), when the following (Ci) is satisfied, the sample is identified as being derived from a subject highly likely to have a 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 low as compared with a predetermined threshold value.

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

6. 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; Based on the concentration of the free fatty acids in the sample obtained in the step (A1), a step (B1) of identifying a sample derived from a subject highly likely to have one or more selected from the group consisting of endometrial cancer, cervical cancer, and precancerous lesions of endometrial cancer and cervical cancer. The method for examining a sample, wherein in the step (B1), when at least one selected from the group consisting of the following (i1), (ii1), and (iii1) is satisfied, the sample is identified as being derived from a subject highly likely to have one or more selected from the group consisting of endometrial cancer, cervical cancer, and precancerous lesions of endometrial cancer and cervical cancer. (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 high as compared with 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 low as compared with a predetermined threshold value; (iii1) The concentration ratio of docosapentaenoic acid to oleic acid is low as compared with a predetermined threshold value.

7. 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; A step (B1) of identifying a sample derived from a subject highly likely to have a gynecological malignant tumor at Stage I or Stage II based on the concentration of the free fatty acid in the sample obtained in the step (A1); A method for examining a sample, wherein in the step (B1), when at least one of the following (i1), (ii1), and (iii1) is satisfied, the sample is identified as being derived from a subject highly likely to have a gynecological malignant tumor at Stage I or Stage II; (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 high as compared with 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 low as compared with a predetermined threshold value; (iii1) The concentration ratio of docosapentaenoic acid to oleic acid is low as compared with a predetermined threshold value.

8. 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, and arachidic 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. The method for examining a sample according to claim 7.

9. The method for examining a sample according to any one of claims 1 to 4 and 6 to 8, wherein the sample is a blood sample.

10. It contains 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, adrenic acid, nervonic acid, lignoceric acid, dihomo-γ-linolenic acid, and docosapentaenoic acid. A kit for examining gynecological cancer and its precancerous lesions, which is used for the purpose of (a) below; (a) Identification of a sample derived from a subject highly likely to have one or more selected from the group consisting of gynecological cancer and its precancerous lesions.

11. The kit for examining gynecological cancer according to claim 10, which is used in the method for examining a sample according to claim 1.

12. It contains 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. A kit for examining gynecological cancer and its precancerous lesions, which is used for at least one purpose selected from the group consisting of (b) to (d) below; (b) Identification of a sample derived from a subject highly likely to have a gynecological cancer with a poor prognosis from a sample of a subject having a gynecological cancer; (c) Evaluation of whether a sample derived from a subject having a gynecological cancer is derived from a subject highly likely to have a gynecological malignancy or a subject highly likely to have a gynecological malignancy; (d) Evaluation of whether a sample identified as being derived from a subject highly likely to have one or more selected from the group consisting of gynecological cancer and gynecological precancerous lesions is derived from a subject highly likely to have a gynecological cancer or a subject highly likely to have a gynecological precancerous lesion.

13. 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, A test kit for uterine body cancer, cervical cancer, and precancerous lesions of uterine body cancer and cervical cancer, which is used for the purpose of (a) below; (a) Identification of a sample derived from a subject highly likely to have one or more selected from the group consisting of uterine body cancer, cervical cancer, and precancerous lesions of uterine body cancer and cervical cancer.

14. 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, A test kit for gynecological cancer and its precancerous lesions, which is used for the purpose of (a) below; (a) Identification of a sample derived from a subject highly likely to have a gynecological malignancy at Stage I or Stage II.

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