Application of ORM1 and / or APOF in the diagnosis and prognosis of cervical cancer and high-grade cervical intraepithelial neoplasia

Abstract

The present invention belongs to the field of disease diagnosis, prognosis and molecular biology technology, and specifically relates to the application of ORM1 and / or APOF in the diagnosis and prognosis of cervical cancer and high-grade cervical intraepithelial neoplasia. The present invention successfully screens relevant biomarkers from the plasma proteins of subjects through DIA and other related technologies. It has been verified by experiments that the area under the curve of ORM1 combined with APOF for predicting healthy subjects and high-grade cervical intraepithelial neoplasia and cervical cancer patients is 0.978. When diagnosing high-grade cervical intraepithelial neoplasia and cervical cancer patients, the AUC can reach 0.982. The high expression of ORM1 is related to lymph node metastasis, clinical stage and poor prognosis of cervical cancer patients. At the same time, it can also be used as a target for screening therapeutic drugs for cervical cancer and high-grade cervical intraepithelial neoplasia, so it has good practical application value.

Description

Technical Field

[0001] The present invention belongs to the technical fields of disease diagnosis, prognosis and molecular biology, and specifically relates to the application of ORM1 and / or APOF in the diagnosis and prognosis of cervical cancer and high-grade cervical intraepithelial neoplasia. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] Cervical cancer is the most common gynecological malignancy in women. According to the latest cancer research data released by the WHO, the number of new cases of cervical cancer in 2020 is about 600,000, and the number of deaths is about 340,000. Although the cause of cervical cancer is relatively clear, it is caused by persistent infection with high-risk human papillomavirus (HPV). However, 90% of HPV-infected patients can automatically clear the HPV virus in their bodies within 2 years, and the process from persistent HPV infection to cervical cancer is slow, suggesting that HPV infection is a necessary condition for the occurrence and development of cervical cancer, but not a sufficient condition. A large number of studies have shown that multiple factors such as intestinal flora imbalance, non-coding RNA and exosome regulation, abnormal methylation and single nucleotide polymorphisms, and destruction of the cervical immune microenvironment are involved in the occurrence and development of cervical cancer.

[0004] Currently, the diagnosis of cervical cancer and cervical intraepithelial neoplasia relies on a three-step approach of HPV / cytology (TCT) screening, colposcopy, and biopsy, which is time-consuming and costly. In most developing countries, there is a shortage of physicians for cytology, colposcopy, and histology diagnosis, resulting in the risk of missed diagnosis and overdiagnosis of cervical cancer and cervical intraepithelial neoplasia. Serological testing is an objective and simple diagnostic method. Currently, serological diagnosis of cervical cancer and cervical intraepithelial neoplasia is relatively scarce. Squamous cell carcinoma antigen (SCC) is a serological detection method for locally advanced cervical cancer, which has poor sensitivity and has no clinical significance in the diagnosis of high-grade cervical intraepithelial neoplasia (HSIL). Summary of the invention

[0005] In view of the deficiencies of the above-mentioned prior art, the inventors, after long-term technical and practical exploration, provide the application of ORM1 and / or APOF in the diagnosis and prognosis of cervical cancer and high-grade cervical intraepithelial neoplasia. The present invention is based on quantitative proteomics technology based on data-independent acquisition (DIA), which provides a comprehensive protein spectrum from plasma, and further reveals the differential proteins between high-grade squamous intraepithelial lesions and cervical cancer as candidate biomarkers. In addition, the present invention verifies the candidate biomarkers of high-grade squamous intraepithelial lesions and cervical cancer patients through subsequent enzyme-linked immunosorbent assay (ELSIA). Based on the above research results, the present invention is completed.

[0006] In order to achieve the above technical objectives, the present invention adopts the following technical solutions:

[0007] The first aspect of the present invention provides the use of a substance for detecting ORM1 and / or APOF in the preparation of a product for diagnosing, detecting, monitoring or predicting the progression of cervical cancer and cervical intraepithelial neoplasia.

[0008] The prediction of the progression of cervical cancer and cervical intraepithelial neoplasia includes but is not limited to the evaluation and analysis of lymph node metastasis, clinical staging and prognosis.

[0009] Wherein, the prognosis at least includes an assessment of the subject's survival period, and the survival period includes progression-free survival and overall survival. The subject may be a cervical cancer patient.

[0010] The second aspect of the present invention provides a product for diagnosing, detecting, monitoring or predicting the progression of cervical cancer and cervical intraepithelial neoplasia, which comprises detecting the expression of ORM1 and / or APOF in a sample based on immunological technology.

[0011] A third aspect of the present invention provides a system for diagnosing, detecting, monitoring or predicting the progression of cervical cancer and cervical intraepithelial neoplasia, the system comprising:

[0012] i) an analysis module, the analysis module comprising a detection substance for determining the expression level of the above-mentioned ORM1 and / or APOF in a test sample of a subject, and;

[0013] ii) an evaluation module, the evaluation module comprising: judging the condition of the subject according to the expression level of the ORM1 and / or APOF determined in i).

[0014] The fourth aspect of the present invention provides the use of the above-mentioned ORM1 and / or APOF as targets for screening drugs for preventing or treating cervical cancer and cervical intraepithelial neoplasia.

[0015] Compared with the existing technical solutions, the above one or more technical solutions have the following beneficial effects:

[0016] The above technical solution successfully screened out ORM1 and APOF from the plasma proteins of the subjects through DIA and other related technologies. It was verified by experiments that the area under the curve (AUC) of ORM1 combined with APOF for predicting healthy subjects and high-grade cervical intraepithelial neoplasia and cervical cancer patients was 0.978. When diagnosing high-grade cervical intraepithelial neoplasia and cervical cancer patients, the AUC can reach 0.982. The high expression of ORM1 is related to lymph node metastasis, clinical stage and poor prognosis of cervical cancer patients. Therefore, ORM1 and APOF can be used as biomarkers for cervical cancer and high-grade cervical intraepithelial neoplasia for the diagnosis and prognosis evaluation of related diseases. At the same time, they can also be used as targets for screening therapeutic drugs for cervical cancer and high-grade cervical intraepithelial neoplasia, so they have good practical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] The accompanying drawings in the specification, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0018] Figure 1 This is the overall project roadmap for DIA sequencing in the embodiments of the present invention.

[0019] Figure 2 The expression of differentially expressed proteins by DIA sequencing in the embodiment of the present invention; wherein, (A) a bar graph of protein quantitative difference results; (B) a volcano graph of differentially expressed proteins between the CC group and the CK group; (C) a volcano graph of differentially expressed proteins between the CC group and the HSIL group; (D) a volcano graph of differentially expressed proteins between the HSIL group and the CK group.

[0020] Figure 3 The quantitative expression diagram of ORM1 (A) and APOF (B) in 60 DIA specimens in the examples of the present invention.

[0021] Figure 4 ELISA validation of the present invention. ELISA was used to detect the expression of ORM1 (A) and APOF (B). (C) ROC of ORM1 and APOF in identifying CK and HSIL / CC; (D) ROC of ORM1 and APOF in distinguishing HSIL and CC. (E and F) PFS comparison between ORM1 / APOF high-expressing cervical cancer patients (20%) and ORM1 / APOF low-expressing cervical cancer patients (80%); (G, H) OS comparison between ORM1 / APOF high-expressing cervical cancer patients (20%) and ORM1 low-expressing cervical cancer patients (80%). DETAILED DESCRIPTION

[0022] It should be noted that the following detailed descriptions are illustrative and are intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present application belongs.

[0023] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present application. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, it indicates the presence of features, steps, operations, devices, components and / or combinations thereof.

[0024] As mentioned above, serological diagnosis of cervical cancer and cervical intraepithelial neoplasia is currently scarce. Squamous cell carcinoma antigen (SCC) is a serological detection method for locally advanced cervical cancer, which has poor sensitivity and has no clinical significance in the diagnosis of high-grade cervical intraepithelial neoplasia (HSIL).

[0025] At present, quantitative proteomics technology based on data independent acquisition is an effective technology for large-scale protein identification and quantification. In the data dependent acquisition (DDA) mode, compared with the traditional mass spectrometry (MS) method, DIA has the advantages of deep proteome coverage, high quantitative repeatability, and high accuracy, and has been applied to biomarker discovery, clinical research, basic exploration and other fields.

[0026] In view of this, the present invention uses the DIA method to provide a comprehensive protein spectrum from plasma, further revealing the differential proteins between high-grade squamous intraepithelial lesions and cervical cancer as candidate biomarkers. In addition, the present invention verifies the candidate biomarkers of high-grade squamous intraepithelial lesions and cervical cancer patients through subsequent enzyme-linked immunosorbent assay (ELSIA).

[0027] Specifically, in a typical embodiment of the present invention, there is provided the use of a substance for detecting ORM1 and / or APOF in the preparation of a product for diagnosing, detecting, monitoring or predicting the progression of cervical cancer and cervical intraepithelial neoplasia.

[0028] The present invention screened and obtained significantly differentially expressed proteins ORM1 and APOF by DIA sequencing technology, and further verified ORM1 and APOF by ELISA method. The results showed that ORM1 combined with or without APOF can become excellent diagnostic indicators for high-grade cervical intraepithelial neoplasia and cervical cancer. Further, by studying the relationship between ORM1 and APOF expression and clinical pathological factors and survival prognosis, the results showed that the expression level of ORM1 in cervical cancer tissue was not related to age, histology, tumor differentiation, clinical stage, tumor size, lymphovascular space invasion (LVSI), deep stromal invasion (DSI), but was related to lymph node metastasis (LN) and clinical stage. However, the above clinical pathological factors were not correlated with the expression of APOF. In terms of survival analysis, the expression of ORM1 and APOF was not associated with progression-free survival (PFS) and overall survival (OS), but when we defined the top 20% as high expression and the remaining 80% as low expression, ORM1 was associated with PFS and OS in these cervical cancer patients.

[0029] Wherein, the ORM1 and / or APOF are of human origin; specifically, derived from the blood of a subject, further from plasma.

[0030] Therefore, the prediction of the progression of cervical cancer and cervical intraepithelial neoplasia includes but is not limited to the evaluation and analysis of lymph node metastasis, clinical staging and prognosis.

[0031] Wherein, the prognosis at least includes an assessment of the subject's survival period, and the survival period includes progression-free survival and overall survival. The subject may be a cervical cancer patient.

[0032] In another specific embodiment of the present invention, a product for diagnosing, detecting, monitoring or predicting the progression of cervical cancer and cervical intraepithelial neoplasia is provided, which comprises a substance for detecting the expression of ORM1 and / or APOF in a sample.

[0033] The ORM1 and / or APOF are of human origin; the substances expressed by ORM1 and / or APOF in the test sample at least include substances for qualitative or quantitative (including semi-quantitative) detection of the above proteins based on immunological techniques, such as Western Blot, ELISA, colloidal gold test strips, protein chips, etc., which are not specifically limited here.

[0034] The sample may be blood of a subject, and further may be plasma.

[0035] The product may be a detection reagent, a detection kit or a biosensor, etc., which is not specifically limited here.

[0036] In another embodiment of the present invention, a system for diagnosing, detecting, monitoring or predicting the progression of cervical cancer and cervical intraepithelial neoplasia is provided, the system comprising:

[0037] i) an analysis module, the analysis module comprising a detection substance for determining the expression level of the above-mentioned ORM1 and / or APOF in a test sample of a subject, and;

[0038] ii) an evaluation module, comprising: analyzing the disease condition of the subject according to the expression level of ORM1 and / or APOF determined in i).

[0039] Wherein, the ORM1 and / or APOF are of human origin. The sample to be tested is the blood of a subject, and further is plasma.

[0040] The detection substances at least include substances for qualitative or quantitative (including semi-quantitative) detection of the above-mentioned proteins based on immunoassay technology, such as Western Blot, ELISA, colloidal gold test strips, protein chips, etc., which are not specifically limited here.

[0041] The analysis of the subject's medical condition at least includes an evaluation of the diagnosis of the subject's cervical cancer and cervical intraepithelial neoplasia, the degree of malignancy (such as lymph node metastasis, clinical stage), and the prognosis of cervical cancer patients (such as overall survival and progression-free survival).

[0042] In another specific embodiment of the present invention, there is provided the use of ORM1 and / or APOF as targets for screening drugs for preventing or treating cervical cancer and cervical intraepithelial neoplasia.

[0043] In another embodiment of the present invention, the effect of a candidate drug on ORM1 and / or APOF before and after use can be used to determine whether the candidate drug can be used to prevent or treat cervical cancer and cervical intraepithelial neoplasia.

[0044] The medicament further comprises at least one pharmaceutically inactive ingredient.

[0045] The inactive ingredients of the drug can be carriers, excipients and diluents commonly used in pharmacy. Moreover, according to the usual methods, the drug can be made into oral preparations, external preparations, suppositories and sterile injection solutions in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, sprays, etc.

[0046] The non-drug active ingredients such as carriers, excipients and diluents that may be included are well known in the art, and those skilled in the art can determine whether they meet clinical standards.

[0047] In another specific embodiment of the present invention, the carrier, excipient and diluent include but are not limited to lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil, etc., and are not specifically limited here.

[0048] The present invention is further described below in conjunction with specific examples, which are only for the purpose of explaining the present invention and are not intended to limit the content thereof. If the specific experimental conditions are not specified in the examples, they are usually carried out according to conventional conditions or the conditions recommended by the sales company; unless otherwise specified in the present invention, they can all be purchased through commercial channels.

[0049] Example

[0050] 1. Research Methods

[0051] (1) DIA identification method

[0052] A total of 125 clinical plasma samples were collected from Qilu Hospital of Shandong University from January 2019 to June 2020, including 30 healthy female volunteers (named CK), 41 HSILs (named HSIL), and 54 cervical cancer patients (named CC), which were divided into a discovery group and a validation group. Among them, 20 CK, 20 HSIL, and 20 CC specimens were analyzed by DIA method for protein changes in these three states, and these 60 specimens belonged to the discovery group. In addition, 30 CK, 41 HSIL, and 54 CC were analyzed by ELSIA to validate the selected proteins. These 125 specimens are the validation group, which includes the discovery group.

[0053] Plasma was collected before breakfast on the second day after hospitalization. Samples were processed immediately according to the standardized protocol recommended by the HUPO plasma proteomics project. Blood was first collected into EDTA tubes (BD), manually inverted 10 times, centrifuged at 4°C 2000r / min for 10 minutes, and then stored at -80°C. At the same time, 10μl of serum from each sample was mixed as a quality control sample. The clinical information of all patients was collected from the hospital's His system. Written informed consent was obtained from all patients. This study was approved by the Ethics Committee of Qilu Hospital of Shandong University (Ethics Number: KYLL-2017-560).

[0054] The overall technical route of the DIA sequencing project is as follows: DIA analysis is completed through sample preparation, protein quantification and electrophoresis, protein enzymatic hydrolysis, LC-MS / MS analysis, etc. First, the samples after equal amounts of merging are used as pool samples for HPRP classification, and LC-MS / MS (QE–HFX_DDA mode) analysis is performed as the database for DIA work. Then, LC-MS / MS (QE–HFX_DDA mode) analysis is performed on each sample separately, and the above database is used for qualitative and quantitative analysis. Finally, bioinformatics analysis tools are used to display and deeply mine the data. The overall project route is as follows: Figure 1 As shown in the figure, it mainly includes two parts: map construction and DIA analysis: protein extraction, peptide enzymatic hydrolysis, chromatographic fractionation, liquid chromatography-tandem mass spectrometry (LC-MS / MS) data DDA acquisition, liquid chromatography-tandem mass spectrometry (LC-MS / MS) data DIA acquisition, library construction, protein DIA data identification and quantitative analysis, differentially expressed protein screening, differentially expressed protein clustering analysis, functional annotation and pathway analysis steps.

[0055] The mass spectrometry method and LC-MS / MS data analysis are as follows:

[0056] DIA analysis was performed using the Easy nLC HPLC system with an ascending flow rate for chromatographic separation. Buffer: Solution A is 0.1% formic acid in water, and solution B is 0.1% formic acid in acetonitrile (84% acetonitrile). The chromatographic column was balanced with 95% solution A. After the sample was injected into the Trap Column, it was gradient separated by passing through the 25cm tip-column of the chromatographic analysis column at a flow rate of 300nl / min. Primary mass spectrometry scanning range: 300-1800m / z, mass spectrometry resolution: 60,000 (@m / z 200), AGC target: 3e6, Maximum IT: 50ms. After each full MS scan, 20 ddMS2 scans (MS2scans) were collected according to the inclusion list, Isolation window: 1.6Th, mass spectrometry resolution: 30,000 (@m / z 200), AGC target: 3e6, Maximum IT: 120ms, MS2 Activation Type: HCD, Normalized collision energy: 27.

[0057] DIA analysis was performed using the Easy nLC HPLC system with an ascending flow rate for chromatographic separation. Buffer: Solution A is 0.1% formic acid in water, and solution B is 0.1% formic acid in acetonitrile (84% acetonitrile). The chromatographic column was balanced with 95% solution A. After the sample was injected into the Trap Column, it was gradient separated by passing through the 25cm tip-column of the chromatographic analysis column at a flow rate of 300nl / min. The primary mass spectrometer scanning range was 300-1650m / z, mass spectrometer resolution: 120,000 (@m / z 200), AGC target: 3e6, Maximum IT: 50ms. MS2 adopted the DIA data acquisition mode, set 30 DIA acquisition windows, mass spectrometer resolution: 15,000 (@m / z200), AGC target: 3e6, Maximum IT: auto, MS2 Activation Type: HCD, Normalized collision energy: 30, Spectral data type: profile.

[0058] LC-MS / MS Data Analysis:

[0059] DDA data were directly imported into Spectronaut software to build a spectral library. The database was downloaded using human_uniprot. The search parameters were set as follows: the enzyme was trypsin, the max miss cleavage site was 1, the fixed modification was Carbamidomethyl (C), the dynamic modification was set to Oxidation (M) and Acetyl (Protein N-term), and the proteins identified by database search must pass the set transition parameter FDR < 1%.

[0060] DIA data were processed using Spectronaut software, and the database was the same as that used for database construction. The software parameters were set as follows: retention time prediction type was set to dynamic iRT, interference on MS2 level correction was enabled, cross run normalization was enabled, and all results were required to pass the set filtering parameter QValue cutoff of 0.01 (equivalent to FDR < 1%).

[0061] (2) ELISA verification method

[0062] ELISA kits were obtained from ELISA Biotechnology Co., Ltd. (Yancheng, Jiangsu, China) and used according to the manufacturer's instructions.

[0063] ORM1 ELSIA Kit No.: MM-61510H1.

[0064] APOF ELISA Kit No.: MM-50834H1.

[0065] 2. Research Results

[0066] (1) Three comparison groups were set up: CC vs CK; CC vs HSIL; HSIL vs CK. By one-way ANOVA, when comparing the three groups, proteins with fold change (FC) ≥ 1.5 or ≤ 0.67 and p value < 0.05 were defined as significantly differentially expressed proteins (DEPs). In this study, a total of 243 DEPs ( Figure 2 A). When analyzing DEPs, we found that compared with CK, the CC group had a total of 81 DEPs, including 51 up-regulated proteins and 30 down-regulated proteins ( Figure 2 A, B). There were 60 DEPs in CC group compared with HSIL group, including 13 up-regulated proteins and 47 down-regulated proteins ( Figure 2 A, C). Compared with the CC group, the HSIL group had 102 DEPs, including 91 up-regulated proteins and 11 down-regulated proteins ( Figure 2 A, D). The above data suggest that it is possible to find suitable biodiagnostic markers for diagnosing HSIL and cervical cancer in our study cohort.

[0067] (2) ORM1 and APOF were identified as two important protein molecules of interest. The DIA expression pattern was as follows Figure 3 shown.

[0068] (3) ELISA validation: To validate the expression patterns of proteins identified by DIA technology, we validated ORM1 and APOF by ELISA, and the results reflected their expression at the plasma protein level. ELISA was performed on 30 healthy volunteers, 41 HSIL and 54 cervical cancer patients. Our results showed that APOF and ORM1 showed almost the same relative expression patterns in DIA and ELISA. Results ( Figure 4A) showed that the plasma expression of ORM1 was the highest in HSIL, followed by cervical cancer, and the lowest in healthy volunteers (1092±7.3 vs 952.2±6.9 ​​vs 859.2±7.4 ng / ml, P<0.001). The APOF in CC group was higher than that in HSIL group (415.4±4.6 vs 402.2±4.5 ng / ml; P=0.0485) and CK (415.4±4.6 vs 364.2±10.8 ng / ml; P<0.001). The APOF in HSIL patients was also higher than that in CK (P=0.0006) ( Figure 4 B). Receiver operating characteristic curve (ROC) Figure 4 C and Figure 4 As shown in Figure D, the area under the curve (AUC) of ORM1 combined with APOF for predicting CK and HSIL+CC was 0.978, and the AUC could reach 0.982 for diagnosing HSIL and CC. The AUCs of ORM1 alone and ORM1+APOF were similar, and the AUC of APOF alone was not ideal. These data suggest that ORM1 combined with or without APOF may be an excellent diagnostic indicator for HSIL and cervical cancer.

[0069] (4) Relationship between ORM1 / APOF expression and clinical pathological factors and survival prognosis

[0070] We performed chi-square test and Kaplan-Meier analysis on the pathological factors related to cervical cancer patients to explore the relationship between ORM1 / APOF expression and clinical pathological factors and survival prognosis. Due to factors such as some patients not undergoing follow-up surgery or being lost to follow-up, a total of 43 cervical cancer patients underwent chi-square test and 35 underwent Kaplan-Meier analysis. The top 50% of cervical cancer patients were defined as high expression, and the rest were low expression. The results are shown in Tables 1 and Figure 4 E, F. We found that the expression level of ORM1 in cervical cancer tissues was not related to age, histology, tumor differentiation, clinical stage, tumor size, lymphovascular space invasion (LVSI), and deep stromal invasion (DSI), but was related to lymph node metastasis (LN) and clinical stage. However, none of the above clinical pathological factors were correlated with the expression of APOF. In terms of survival analysis, the expression of ORM1 and APOF was not correlated with progression-free survival (PFS) and overall survival (OS), but when we defined the top 20% of expression as high expression and the remaining 80% as low expression, ORM1 was associated with PFS and OS in these cervical cancer patients ( Figure 4 E, G).

[0071] Table 1 Relationship between ORM1 / APOF expression and clinical pathological factors

[0072]

[0073]

[0074] In summary, the area under the curve (AUC) of ORM1 combined with APOF for predicting CK and HSIL+CC was 0.978, and the AUC could reach 0.982 when diagnosing HSIL and CC. High expression of ORM1 was associated with lymph node metastasis, clinical stage and poor prognosis in patients with cervical cancer. ORM1 and APOF can be used as biomarkers for cervical cancer and high-grade cervical intraepithelial neoplasia.

[0075] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. Use of a substance for detecting a combination of ORM1 and APOF in the preparation of a product for diagnosing or predicting the progression of cervical cancer and cervical intraepithelial neoplasia, wherein the combination of ORM1 and APOF is derived from the plasma of a subject; The applications include differentiating or identifying cervical cancer and cervical intraepithelial neoplasia; The prediction of the progress of cervical cancer and cervical intraepithelial neoplasia includes the evaluation and analysis of lymph node metastasis, clinical stage and prognosis; in, The prognosis at least includes an assessment of the subject's survival, which includes progression-free survival and overall survival; the subject is a cervical cancer patient.

2. The use according to claim 1, characterized in that The ORM1 and APOF combination is of human origin.

3. A system for diagnosing or predicting the progression of cervical cancer and cervical intraepithelial neoplasia, characterized in that: The system comprises: i) an analysis module, the analysis module comprising a detection substance for determining the expression level of the combination of ORM1 and APOF selected from the above in a test sample of a subject, and; ii) an evaluation module, the evaluation module comprising: analyzing the disease condition of the subject according to the expression level of the combination of ORM1 and APOF determined in i); The combination of ORM1 and APOF is of human origin; the sample to be tested is the plasma of a subject.

4. The system according to claim 3, characterized in that The detection substance at least comprises a substance for qualitative or quantitative detection of the above-mentioned protein based on immunological technology; Analyzing the subject's medical condition at least includes evaluating the diagnosis, malignancy, and prognosis of cervical cancer and cervical intraepithelial neoplasia of the subject; The malignancy degree includes lymph node metastasis and clinical stage; The prognoses included overall survival and progression-free survival.

5. Use of ORM1 and APOF combination as targets for screening drugs for preventing or treating cervical cancer and cervical intraepithelial neoplasia.

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