Application of an ADIRF detection reagent in the preparation of ovarian cancer drug resistance diagnostic reagent

Abstract

The present invention discloses an application of an ADIRF detection reagent in the preparation of an ovarian cancer drug resistance diagnostic reagent. Ovarian cancer peritoneal metastasis is the most common form of metastasis of ovarian cancer and is also the most important link in the progression of the disease. Platinum-based chemotherapy is the main means of eliminating ovarian cancer cells with peritoneal metastasis, but it does not achieve the desired effect for drug-resistant patients. Therefore, the diagnosis of drug resistance is an important basis for clinical decision-making. The discovery and application of new markers contribute to the development of diagnostic methods. The present invention finds through research that ADIRF can be used as a diagnostic marker for ovarian cancer peritoneal metastasis resistance. Therefore, the present invention provides a new application of a detection reagent using the new marker ADIRF to prepare an ovarian cancer peritoneal metastasis resistance diagnostic reagent. The present invention contributes to the development of a method for diagnosing ovarian cancer peritoneal metastasis resistance.

Description

Technical Field

[0001] The present invention relates to the field of clinical testing and diagnosis, and in particular to application of an ADIRF detection reagent in the preparation of an ovarian cancer drug resistance diagnosis reagent. Background Art

[0002] Chemotherapy is the main treatment for ovarian cancer, but chemotherapy-resistant ovarian cancer cells are resistant to chemotherapy. Therefore, determining the drug resistance of ovarian cancer is an important basis for clinical decision-making.

[0003] The properties and capabilities of cells are closely related to the characteristics of their protein expression. The functions of proteins expressed in cells determine the properties and capabilities of cells. Therefore, the diagnosis of drug resistance of ovarian cancer cells requires appropriate protein markers.

[0004] In addition, the combination of markers can effectively improve the accuracy of the diagnostic method, that is, improve the sensitivity and specificity. Therefore, the identification of new and effective markers has a promoting effect on the diagnosis of ovarian cancer drug resistance.

[0005] Based on the above research significance, the present invention uses proteomics research technology to compare the protein expression profiles of ovarian cancer peritoneal metastasis tissues before and after neoadjuvant chemotherapy, and finds that ADIRF is highly expressed in residual tissues after neoadjuvant chemotherapy and in peritoneal metastasis tissues of ovarian cancer without response before neoadjuvant chemotherapy. The AUC value for distinguishing non-responsive patients is 0.8126. These research results show that ADIRF is a new marker for judging ovarian cancer drug resistance. Therefore, the present invention provides a method for preparing an ovarian cancer drug resistance diagnostic reagent based on a new marker, which is useful for ovarian cancer drug resistance diagnosis. Summary of the invention

[0006] In view of this, the object of the present invention is to provide an application of an ADIRF detection reagent in the preparation of an ovarian cancer drug resistance diagnostic reagent.

[0007] In order to achieve the above object, the present invention provides the following technical solutions:

[0008] Application of an ADIRF detection reagent in the preparation of an ovarian cancer drug resistance diagnostic reagent.

[0009] Preferably, the gene ID of the ADIRF is 10974, and the Uniprot database ID is Q15874.

[0010] Preferably, the detection reagent of the present invention is used to detect the protein expression level of ADIRF in target cells.

[0011] Preferably, the ovarian cancer refers to primary, peritoneal metastasis and other metastatic ovarian cancers.

[0012] Preferably, in the present invention, the drug resistance refers to drug resistance based on platinum-based drug treatment.

[0013] Preferably, the drug resistance of the present invention refers to the drug resistance of neoadjuvant chemotherapy for peritoneal metastasis of ovarian cancer.

[0014] Preferably, the drug resistance of the present invention refers to conventional chemotherapy resistance of ovarian cancer peritoneal metastasis.

[0015] Preferably, the drug resistance of the present invention refers to the ability of cancer cells to resist the inhibitory effect of treatment methods.

[0016] Preferably, the drug resistance diagnostic reagent of the present invention refers to a reagent capable of diagnosing whether the ovarian cancer cells carried by the patient have drug resistance.

[0017] The benefits of the present invention are as follows: the present invention develops a new application of a reagent for identifying drug resistance of ovarian cancer peritoneal metastasis based on the new marker ADIRF. A method for preparing a reagent for identifying drug resistance of ovarian cancer peritoneal metastasis based on the ADIRF protein detection reagent is also provided. ADIRF can be used as a single marker to identify neoadjuvant chemotherapy-resistant ovarian cancer peritoneal metastasis, and can also be used in combination with other markers. The use provided by the present invention can improve the accuracy of diagnosis of drug resistance of ovarian cancer peritoneal metastasis. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to make the purpose, technical solution and beneficial effects of the present invention clearer, the present invention provides the following drawings for illustration:

[0019] Figure 1 To study strategies for the discovery of chemotherapy-related proteins;

[0020] Figure 2 The difference in proteomic profiles of peritoneal metastasis tissues from ovarian cancer before and after neoadjuvant chemotherapy;

[0021] Figure 3 is the relative mass spectrometry detection signal value of the protein level of ADIRF in peritoneal metastasis tissue of ovarian cancer before and after neoadjuvant chemotherapy;

[0022] Figure 4 Immunohistochemical scoring of ADIRF protein expression in peritoneal metastasis tissues of ovarian cancer before and after neoadjuvant chemotherapy;

[0023] Figure 5 This study is to study the correlation between ADIRF protein expression level and the response to neoadjuvant chemotherapy for peritoneal metastasis of ovarian cancer.

[0024] Figure 6 To evaluate the accuracy of ADIRF as a drug resistance marker for diagnosing peritoneal metastasis of ovarian cancer. DETAILED DESCRIPTION

[0025] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it, but the embodiments are not intended to limit the present invention.

[0026] Example 1: Study on ADIRF as a drug resistance marker for ovarian cancer

[0027] Neoadjuvant chemotherapy is a treatment that uses chemotherapy before surgery to achieve a certain degree of tumor reduction effect to pave the way for surgery. Taking advantage of the fact that residual tumor tissue after neoadjuvant chemotherapy is enriched with resistant tumor cells, this study discovered ovarian cancer resistance-related markers by comparing the differences in protein profiles of ovarian cancer peritoneal metastasis samples after neoadjuvant chemotherapy. The proteomics technology used in this study can simultaneously detect the expression of thousands of proteins in target samples, thereby drawing a proteomics map. Research ideas such as Figure 1 As shown in the figure, the study collected 3 cases of peritoneal metastasis tissues of ovarian cancer before (Pre-NACT) and after (Post-NACT) neoadjuvant chemotherapy (NACT), and used TMT proteomics technology to detect the protein expression levels at the proteomic level in the tissues, and then compared and screened the differentially expressed proteins in the Post-NACT samples ( Figure 2 Among them, ADIRF was one of the proteins with significantly increased expression in Post-NACT samples ( Figure 3 ). This result indicates that ADIRF is a candidate protein in the screening study of ovarian cancer resistance-related proteins, and subsequent verification is needed to prove its nature as a marker of ovarian cancer resistance.

[0028] For the methods of sampling ovarian cancer samples before and after neoadjuvant chemotherapy and detecting them by TMT proteomics technology in this example, please refer to Shen Y, Ren Y, Chen K, Cen Y, Zhang B, Lu W, Xu J. The impact of neoadjuvant chemotherapy on the tumor microenvironment in advanced high-grade serous carcinoma. Oncogenesis. 2022 Jul 30;11(1):43. doi: 10.1038 / s41389-022-00419-1. PMID: 35907904; PMCID: PMC9338965. and McDermott JE, Arshad OA, PetyukVA, Fu Y, Gritsenko MA, Clauss TR, Moore RJ, Schepmoes AA, Zhao R, Monroe ME, Schnaubelt M, Tsai CF, Payne SH, Huang C, Wang LB, Foltz S, Wyczalkowski M, Wu Y, Song E, Brewer MA, Thiagarajan M, Kinsinger CR, Robles AI, Boja ES, Rodriguez H, Chan DW, Zhang B, Zhang Z, Ding L, Smith RD, Liu T, Rodland KD; Clinical Proteomic Tumor Analysis Consortium. Proteogenomic Characterization of Ovarian HGSC Implicates Mitotic Kinases, Replication Stress in ObservedChromosomal Instability. Cell Rep Med. 2020 Apr 21;1(1):100004. doi: 10.1016 / j.xcrm.2020.100004. Epub 2020 Apr 10. Erratum in: Cell Rep Med. 2020 Aug 25;1(5): PMID: 32529193; PMCID: PMC7289043.

[0029] Example 2: Immunohistochemical analysis of the protein expression level of ADIRF in peritoneal metastasis tissues of ovarian cancer before and after neoadjuvant chemotherapy

[0030] To further verify the differential expression of ADIRF in peritoneal metastasis tissues of ovarian cancer after neoadjuvant chemotherapy, this study conducted a validation study in 53 pairs of the same tissues before and after chemotherapy. The validation method used a more accurate immunohistochemical detection method to analyze the samples one by one. The test results were expressed as the immunoreactive score. Figure 4 As shown in the results, the ADIRF in peritoneal metastasis tissues of ovarian cancer after neoadjuvant chemotherapy was significantly increased compared with that before neoadjuvant chemotherapy. This result certainly suggests the correlation between ADIRF and chemotherapy resistance in ovarian cancer.

[0031] For the immunohistochemistry operation and result interpretation method in this example, please refer to Fedchenko N, Reifenrath J. Different approaches for interpretation and reporting of immunohistochemistry analysis results in the bone tissue - a review. DiagnPathol. 2014 Nov 29;9:221. doi: 10.1186 / s13000-014-0221-9. PMID: 25432701;PMCID: PMC4260254.

[0032] Example 3. Study on the correlation between ADIRF protein expression level and response to neoadjuvant chemotherapy for ovarian cancer peritoneal metastasis

[0033] According to the degree of response of the tumor tissue carried by the patient after chemotherapy to neoadjuvant chemotherapy, it can be divided into complete response, partial response and no response. In order to further verify the correlation between ADIRF and the resistance of neoadjuvant chemotherapy in peritoneal metastasis of ovarian cancer, this study collected 67 pre-chemotherapy samples with complete or partial response and 100 no response, and detected the ADIRF protein expression level of each sample by immunohistochemistry. The correlation between the protein expression level of ADIRF and the degree of response to neoadjuvant chemotherapy in peritoneal metastasis of ovarian cancer was statistically analyzed by chi-square analysis. The results are as follows Figure 5 The results showed that the protein expression level of ADIRF was negatively correlated with the degree of response. This result shows that the protein expression level of ADIRF has the ability to be used as an indicator to predict the therapeutic effect before neoadjuvant chemotherapy.

[0034] For the immunohistochemistry operation and result interpretation method in this example, please refer to Fedchenko N, Reifenrath J. Different approaches for interpretation and reporting of immunohistochemistry analysis results in the bone tissue - a review. DiagnPathol. 2014 Nov 29;9:221. doi: 10.1186 / s13000-014-0221-9. PMID: 25432701;PMCID: PMC4260254. For the evaluation method of neoadjuvant chemotherapy response in this example, see Ramspott JP, BaertT, MacKintosh ML, Traut A, Ataseven B, Bommert M, Heitz F, Plett H, SchneiderS, Waltering KU, Heikaus S, Harter P, du Bois A. Response evaluation afterneoadjuvant therapy: evaluation of chemotherapy response score andserological and / or radiological assessment of response in ovarian cancerpatients. Arch Gynecol Obstet. 2021 Oct;304(4):1021-1032. doi: 10.1007 / s00404-021-06020-y. Epub 2021 Mar 4. PMID: 33661392.

[0035] Example 4: Evaluation of the accuracy of ADIRF as a drug resistance marker for diagnosing peritoneal metastasis of ovarian cancer

[0036] This study further evaluated the accuracy of ADIRF as a marker for judging the drug resistance of ovarian cancer peritoneal metastasis by AUC value. Based on the test results in Example 3, ROC curves were drawn and AUC values ​​were calculated to evaluate the accuracy of ADIRF in identifying patients with non-responsive ovarian cancer peritoneal metastasis before treatment. Figure 6As shown in the figure, the AUC value is 0.8126, indicating that when the immunohistochemical score is set as a threshold of 5, the sensitivity of ADIRF as a marker to distinguish non-responsive patients is 77%, and the specificity is 76.12%. The above results show that ADIRF has the ability to be used as a marker to evaluate the drug resistance of ovarian cancer, and the accuracy of diagnosis can be improved by combining it with more markers.

[0037] The above-described embodiments are only preferred embodiments for fully illustrating the present invention, and the protection scope of the present invention is not limited thereto. Equivalent substitutions or changes made by those skilled in the art based on the present invention are within the protection scope of the present invention. The protection scope of the present invention shall be subject to the claims.

Claims

1. An application of an ADIRF detection reagent in the preparation of an ovarian cancer drug resistance diagnostic reagent, characterized in that: The drug resistance refers to the resistance to neoadjuvant chemotherapy for peritoneal metastasis of ovarian cancer based on platinum drugs.

2. The use according to claim 1, characterized in that: The gene ID of the ADIRF is 10974, and the Uniprot database ID is Q15874.

3. The use according to claim 1, characterized in that: The detection reagent is used to detect the protein expression level of ADIRF in target cells.

4. The use according to claim 1, characterized in that: The ovarian cancer refers to primary, peritoneal metastasis and other metastatic ovarian cancers.

5. The use according to claim 1, characterized in that: Drug resistance refers to the ability of cancer cells to resist the inhibitory effects of therapeutic methods.

6. The use according to claim 1, characterized in that: Drug resistance diagnostic reagents refer to reagents that can diagnose whether ovarian cancer cells carried by a patient are drug resistant.

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