Endometrial cancer conservation treatment resistance markers, products and applications

Abstract

The present invention relates to drug resistance markers, products and applications for conservation treatment of endometrial cancer, and to the technical field of endometrial cancer fertility maintenance treatment. The drug resistance markers for conservation treatment of endometrial cancer include at least 4 of DYNLT1 protein, HSD17B12 protein, EEF1E1 protein, ILVBL protein, SRPK1 protein and NUDT5 protein. The protein markers for conservation treatment of endometrial cancer obtained by screening and the expression levels of the 6 protein markers can be used to predict the treatment response of patients with endometrial cancer before conservation treatment, thereby effectively screening the drug-resistant population, and optimizing the treatment plan in a targeted manner, thereby providing the patient with a benefit rate.

Description

Technical Field

[0001] The present invention relates to the technical field of endometrial cancer fertility-maintaining treatment, and in particular to endometrial cancer conservation treatment drug resistance markers, products and applications. Background Art

[0002] Endometrial carcinoma (EC) is one of the three most common malignant tumors of the female reproductive system. The incidence of endometrial cancer is closely related to risk factors such as obesity and diabetes. In recent years, the incidence of endometrial cancer has shown a trend of increasing year by year and becoming younger, accounting for about 7% of the total number of female cancers and more than 30% of female reproductive system malignant tumors, which is close to or even exceeds the incidence of cervical cancer.

[0003] In recent years, the number of patients with early-onset endometrial cancer has been increasing. These women of childbearing age often have a strong desire for fertility and often choose fertility preservation treatment based on medroxyprogesterone acetate (MPA) to preserve the patient's reproductive function. Therefore, conservation treatment has received increasing attention. Although the highly effective progestin-based regimen has achieved satisfactory results, more than 30% of patients are insensitive to progestin treatment and become refractory patients. The clinical problem of progestin-insensitive patients needs to be solved urgently: how to screen out patients who are insensitive to progestin treatment before the start of treatment in order to develop a treatment plan suitable for these patients.

[0004] Over the past few decades, researchers have found that there is great consistency in the morphological and molecular characteristics of endometrial cancer. With the in-depth study of the molecular mechanisms of tumor pathogenesis and the maturity of bioinformatics technologies such as second-generation sequencing, molecular classification of endometrial cancer has been proposed to assist clinicians in predicting patient prognosis and targeted drugs. However, previous studies only included patients with endometrial cancer who underwent surgical treatment, and the molecular landscape of patients receiving conservative treatment is still blank, and the molecular characteristics of conservative patients with different treatment responses are still unclear. In view of this, the present invention provides endometrial cancer conservative treatment resistance markers, products and applications. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide markers, products and applications of drug resistance in the conservation treatment of endometrial cancer. The purpose is to use the DYNLT1, HSD17B12, EEF1E1, ILVBL, SRPK1 and NUDT5 protein markers obtained by screening to prepare products for predicting the response to conservation treatment of endometrial cancer, so as to judge the drug resistance of patients undergoing conservation treatment of endometrial cancer.

[0006] The present invention solves the above technical problems. The first aspect is to provide endometrial cancer conservation treatment resistance markers, including at least four of DYNLT1 protein, HSD17B12 protein, EEF1E1 protein, ILVBL protein, SRPK1 protein and NUDT5 protein.

[0007] The present invention screened markers of resistance to conservative therapy for endometrial cancer based on mass spectrometry-based proteomic technology, and verified them using immunohistochemical staining. Six protein molecules (DYNLT1, HSD17B12, EEF1E1, ILVBL, SRPK1 and NUDT5) were constructed for predicting resistance to conservative therapy in patients with endometrial cancer.

[0008] The beneficial effects of the present invention are:

[0009] (1) The present invention obtains protein markers of resistance to conservative treatment of endometrial cancer through screening, and by detecting the expression levels of 6 protein markers, it can predict the treatment response of patients with endometrial cancer before they undergo conservative treatment, thereby effectively screening the drug-resistant population and optimizing their treatment plans in a targeted manner, thereby improving the patient's benefit rate.

[0010] (2) The biomarkers explored in the present invention suggest that using DYNLT1, HSD17B12, EEF1E1, ILVBL, SRPK1 and NUDT5 proteins as targets can reverse progesterone therapy resistance, and therefore have broad clinical application prospects.

[0011] The second aspect of the present invention provides the use of endometrial cancer conservation therapy resistance markers, and the reagents for detecting the above-mentioned endometrial cancer conservation therapy resistance markers are used to prepare endometrial cancer conservation therapy resistance products.

[0012] The beneficial effect of adopting the above scheme is: by using reagents for detecting the expression levels of 6 protein markers (DYNLT1, HSD17B12, EEF1E1, ILVBL, SRPK1 and NUDT5) in the preparation of endometrial cancer conservation treatment resistance products, a prediction system for conservation treatment resistance of endometrial cancer patients based on 6 proteins was successfully constructed, which can predict the treatment response of endometrial cancer patients before conservation treatment, and thus effectively screen the resistant population.

[0013] Furthermore, by quantitatively detecting the endometrial cancer conservation treatment drug resistance marker, the drug resistance of patients undergoing endometrial cancer conservation treatment can be determined.

[0014] Furthermore, the drug for the conservation treatment of endometrial cancer is progesterone.

[0015] Furthermore, the drug for the conservation treatment of endometrial cancer is medroxyprogesterone acetate.

[0016] In the present invention, "conservation therapy" is a treatment strategy for patients with early endometrial cancer. The goal of conservation therapy is to preserve the patient's uterus and reproductive function as much as possible, while effectively controlling the development and spread of cancer. Its main method is based on progestin therapy: oral high-efficiency progesterone is currently the most commonly used conservation therapy method. Progesterone inhibits the proliferation of cancer cells by changing the growth and differentiation of endometrial cells, thereby achieving the therapeutic effect. Conservation therapy resistance refers to whether the patient has disease progression within 6 months or no relief within 12 months after receiving oral medroxyprogesterone acetate (MPA) treatment. The conservation therapy resistance state of endometrial cancer can be predicted by the expression level of marker proteins in the patient's tumor tissue before treatment.

[0017] Furthermore, the reagent for detecting the endometrial cancer conservation therapy resistance marker includes a substance that specifically binds to the DYNLT1 protein, the HSD17B12 protein, the EEF1E1 protein, the ILVBL protein, the SRPK1 protein and the NUDT5 protein.

[0018] Furthermore, the substance is an antibody or an antibody fragment.

[0019] The substances for detecting DYNLT1, HSD17B12, EEF1E1, ILVBL, SRPK1 and NUDT5 of the present invention include antibodies or fragments thereof that specifically bind to the target protein. Antibodies or fragments thereof of any structure, size, immunoglobulin class, origin, etc. can be used as long as it binds to the target protein. The antibodies or fragments thereof of the present invention can be monoclonal or polyclonal. An antibody fragment refers to a portion of an antibody (partial fragment) or a peptide containing a portion of an antibody that retains the binding activity of the antibody to the antigen. Antibody fragments may include F(ab')2, Fab', Fab, single-chain Fv (scFv), disulfide-bonded Fv (dsFv) or its polymer, dimerized V region (diabody), or a peptide containing CDR. The substances for detecting DYNLT1, HSD17B12, EEF1E1, ILVBL, SRPK1 and NUDT5 proteins of the present invention may include an isolated nucleic acid encoding an amino acid sequence of an antibody or an antibody fragment, a vector containing the nucleic acid, and a cell carrying the vector.

[0020] The above antibodies can be obtained by methods well known to those skilled in the art. For example, a polypeptide or a mammalian cell expression vector incorporating a polynucleotide encoding a target protein can be prepared as an antigen, retaining the entire or part of the target protein. After the animal is immunized with the antigen, the immune cells can be collected and fused with myeloma cells to obtain a hybridoma. The antibodies can then be collected from the hybridoma culture. Finally, DYNLT1, HSD17B12, EEF1E1, ILVBL, SRPK1 and NUDT5 proteins or parts thereof can be used as antigens for antigen-specific purification to obtain monoclonal antibodies against these proteins. The method for preparing polyclonal antibodies is as follows: immunize the animal with the same antigen as described above, collect a blood sample from the immunized animal, separate the serum, and then use the above antigen to purify the serum antigen-specifically. Antibody fragments can be obtained by enzymatic treatment of the antibodies obtained or by using the obtained antibody sequence information.

[0021] The third aspect of the present invention provides a product for detecting drug resistance markers for conservation therapy of endometrial cancer, the product comprising a reagent for detecting the above-mentioned drug resistance markers for conservation therapy of endometrial cancer.

[0022] The products of the present invention for detecting DYNLT1, HSD17B12, EEF1E1, ILVBL, SRPK1 and NUDT5 proteins can be performed and functioned based on known methods using the above antibodies or antibody fragments, including ELISA, Western blotting, immunohistochemical staining, etc. The products of DYNLT1, HSD17B12, EEF1E1, ILVBL, SRPK1 and NUDT5 proteins can also be quantitatively determined based on mass spectrometry technology, such as protein non-labeling quantitative technology (4D-Label Free) and TMT (Tandem MassTag) labeling quantitative technology.

[0023] The beneficial effect of adopting the above scheme is that the present invention constructs a detection product for predicting conservation resistance, which only requires collecting the patient's tumor tissue to conduct the test according to the manual and evaluate whether the patient will develop drug resistance based on the staining results, which has the advantages of clinical accessibility and accuracy.

[0024] Furthermore, the product includes detection reagents, test kits, chips, test strips or high-throughput sequencing platforms.

[0025] Furthermore, the high-throughput sequencing platform includes a Tims-TOF mass spectrometer.

[0026] The fourth aspect of the present invention provides a method for screening endometrial cancer conservation treatment resistance markers, comprising the following steps:

[0027] (1) Obtain tumor tissue samples from patients who have received conservation therapy;

[0028] (2) Lysing the patient's tissue, extracting the protein therein, and digesting it with pancreatic enzymes to obtain cleaved peptides;

[0029] (3) Use label-free technology to perform high-throughput sequencing of peptides based on the Tims-TOF platform to obtain peptide sequencing results;

[0030] (4) using MaxQuant software to search the obtained peptide sequencing results to obtain high-throughput quantitative results of each protein;

[0031] (5) Patients with different treatment responses were grouped, the quantitative levels of each protein were analyzed between the groups, and the Wilcoxon method was used to test statistical significance;

[0032] (6) Proteins that are significantly upregulated in the conservation resistance group can be considered to be related to conservation resistance.

[0033] The above samples came from tumor tissues of patients receiving conservation treatment before treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 The present invention is based on the proteome to screen conservation-related markers for endometrial cancer; A is a protein volcano plot of conservation-related groups; B is a mesenchymal transition pathway in which proteins of conservation-resistant patients are significantly enriched; C and D are survival analyses of 33 resistance-related proteins in 6 external data sets; P<0.1 of the Log-rank test in at least three data sets; hazard ratio>2;

[0035] Figure 2 The present invention is the verification of the conservation resistance-related markers of endometrial cancer and the construction of the conservation resistance system; wherein A is the representative image of EEF1E1, ILVBL, SRPK1 and NUDT5 immunohistochemical staining in conservation-sensitive and conservation-resistant patients; B is the immunohistochemical staining score of EEF1E1, ILVBL, SRPK1 and NUDT5 in conservation-sensitive and conservation-resistant patients; C is the correlation between EEF1E1, ILVBL, SRPK1 and NUDT5, *P<0.05; **P<0.01, ***P<0.001. DETAILED DESCRIPTION

[0036] The principles and features of the present invention are described below, and the examples are only used to explain the present invention and are not used to limit the scope of the present invention. If no specific technology or conditions are specified in the embodiments, the technology or conditions described in the literature in this field or the product instructions are used. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased through regular channels.

[0037] Example 1

[0038] The method for screening drug resistance-related markers provided by the present invention comprises the following steps:

[0039] a) Sample collection

[0040] This study complies with the ethical standards of the Declaration of Helsinki and the ethics approval numbers are [TJ-IRB20211005] and [No. 2021-181]. Written informed consent was obtained from each patient before any specific study was conducted.

[0041] The present invention screened 62 patients with endometrial cancer who received conservation treatment from January 2015 to July 2020. Histological sections of tumors / adjacent tissues were taken, and hematoxylin-eosin staining (HE) was used to determine the pathological grade of tumor tissue and the proportion of tumor cancer. Each tumor / adjacent sample was examined by 2 pathology experts, and the sample quality was confirmed according to the following criteria: (1) endometrioid endometrial cancer defined by histopathology; (2) the tumor cell rate (tumor purity) after removing the tumor tissue on the section under the naked eye was >90%; (3) there were no tumor cells in the adjacent paracancerous tissue. All cases were staged according to the International Federation of Gynecology and Obstetrics (FIGO) diagnostic criteria (2018). Other inclusion criteria were: the patient had not received any treatment before admission, had no other malignant tumors, had no other hormone-related diseases, and the patient had complete clinical information.

[0042] b) FFPE sample protein extraction and trypsin digestion

[0043] FFPE specimens were scraped from tissue sections and transferred to 1.5 mL conical tubes, and paraffin was removed using a diluent of ethanol and water. Specifically, the tissue was dewaxed with xylene, and then four volumes of lysis buffer were added to the tissue, which was a mixture of 1% SDC (sodium deoxycholate, purchased from Sigma-Aldrich, catalog number D5670) and 1% protease inhibitor (purchased from ServiceBio, catalog number G2006-250UL), and sonicated on ice for 3 minutes. The remaining debris was removed by centrifugation at 12000g for 10 minutes at 4°C, and the supernatant was finally collected and the protein concentration was determined using a BCA kit (purchased from Beyotime, catalog number P0009).

[0044] The extracted protein sample was added to 1% SDC buffer, 5mM dithiothreitol (purchased from Sigma, product number 43816) was added and incubated at 56°C for 30 minutes, and 11mM iodoacetamide (purchased from Sigma, product number A3221) was added for alkylation for 15 minutes at room temperature in the dark. Then, the first overnight digestion was performed with a trypsin (purchased from Sigma, product number T6567) / protein mass ratio of 1:50, and the second 4-hour digestion was performed with a trypsin / protein mass ratio of 1:100. The enzymatic peptides were acidified to pH 2-3 with 10% TFA (trifluoroacetic acid, purchased from Sigma, product number T6508), centrifuged at 12000g for 10 minutes at room temperature, and the supernatant was transferred to a new EP tube. After the SPE column was activated with methanol, it was equilibrated with 0.1% TFA, and the acidified peptide solution was loaded onto the SPE column. It was then desalted with 0.1% TFA, and finally the peptide was eluted with 80% ACN (acetonitrile, purchased from Sigma, catalog number 34851), and then vacuum concentrated and stored at -80°C for future use.

[0045] c) Liquid chromatography-mass spectrometry / mass spectrometry (LC-MS / MS) analysis and protein quantification

[0046] The peptides were dissolved in liquid chromatography mobile phase A and separated using the NanoElute ultra-high performance liquid chromatography system. Mobile phase A was an aqueous solution containing 0.1% formic acid and 2% acetonitrile; mobile phase B was a solution containing 0.1% formic acid and 100% acetonitrile. A 90-minute liquid phase gradient was used, and the flow rate was maintained at 450 nL / min.

[0047] The peptides were separated by ultra-high performance liquid phase system and injected into the Capillary ion source for ionization and then analyzed by timsTOF Pro mass spectrometer. The ion source voltage was set to 1.65kV, and the peptide parent ion and its secondary fragments were detected and analyzed using high-resolution TOF. The secondary mass spectrometer scan range was set to 100-1700. The data acquisition mode used parallel accumulation serial fragmentation (PASEF) mode. After one primary mass spectrum was acquired, 10 secondary spectra were acquired in PASEF mode.

[0048] The secondary mass spectrometry data were retrieved using Maxquant (v1.6.15.0). Search parameter settings: the database was Homo_sapiens_9606_SP_20210721.fasta (20387 sequences), a reverse library was added to calculate the false positive rate (FDR) caused by random matching, and a common contamination library was added to the database to eliminate the impact of contaminating proteins in the identification results; the enzyme cutting method was set to Trypsin / P; the number of missed cutting sites was set to 2; the mass error tolerance of the secondary fragment ion was 20ppm. The cysteine ​​alkylation Carbamidomethyl (C) was set as a fixed modification, and the variable modification was the oxidation of methionine and the acetylation of the protein N-terminus. The FDR for protein identification and PSM identification was set to 1%.

[0049] A total of 95,292 peptides and 7,409 proteins were identified using a permutation-based false discovery rate (FDR < 1%) to correct for multiple hypothesis testing. The data were then subjected to the K nearest neighbor (KNN) algorithm to fill in null values ​​and logarithmically transformed for subsequent analysis.

[0050] d) Proteomic data preprocessing and normalization

[0051] Proteomics analysis was performed in the MaxQuant environment using a permutation-based false discovery rate (FDR) to correct for multiple hypothesis testing (FDR < 1%; Appendix Figure 2 ). The quality of the proteomics data was ensured by further analysis of peptide length, number of peptides, protein molecular weight, and protein coverage. The proteomics data were log2 transformed, and the k-Nearest Neighbor (KNN) algorithm was used to fill missing values ​​for proteins quantified in at least half of the samples. This computational method was implemented in the DreamAI R package.

[0052] e) Screening process of proteins related to drug resistance

[0053] The patients were divided into two groups based on the response to conservation therapy: conservation therapy resistant (n=30) and conservation therapy sensitive (n=32). The proteomic data obtained after the above filling were standardized and normalized to ensure the accuracy and comparability of the data. In order to identify proteins with significant differences between different groups, the present invention uses the Wilcoxon statistical test to explore proteins with statistically significant differences.

[0054] f) Screening results of proteins related to drug resistance

[0055] The results of differential analysis showed that 33 proteins were significantly upregulated in patients with drug resistance to conservation therapy (Wilcoxon's sp < 0.05, Fold Change > 1.5) (results as shown in Figure 1 AB).

[0056] In order to further screen and verify the association of these molecules with the response to conservation therapy, the present invention performed a Log Rank test on the correlation between 33 candidate proteins and endocrine therapy response in 6 public data sets of breast cancer treated with tamoxifen (GSE1378, GSE1379, GSE6532, GSE9195, GSE12093, GSE17705). The present invention found that 6 genes (EEF1E1, DYNLT1, ILVBL, HSD17B12, NUDT5 and SRPK1) among the 33 candidate genes were associated with endocrine therapy resistance (Log Rank's p<0.1; Hazard Ratio>2). The 6 proteins obtained are proteins associated with endometrial cancer resistance (results as shown in Figure 1 CD).

[0057] Example 2

[0058] The process of verifying drug resistance-related markers and constructing a conservation drug resistance prediction system provided by the present invention is as follows:

[0059] a) Sample collection

[0060] The sample collection method is the same as that in Specific Example 1.

[0061] b) Immunohistochemistry (IHC) staining

[0062] 1. Select the paraffin tissue sections that need to be stained with immunohistochemistry, and soak them in environmentally friendly tissue dewaxing solution (environmentally friendly dewaxing transparent solution, purchased from Sevier Biotechnology, item number G1128-1L) for 2 hours.

[0063] 2. Take out the tissue sections and immerse them in new environmentally friendly tissue dewaxing solution for 20 minutes.

[0064] 3. Take out the tissue slices and hydrate them through different gradient concentrations of ethanol, the concentrations of ethanol are 95%, 85%, and 75% respectively. Soak the tissue slices in each concentration of ethanol for 10 minutes.

[0065] 4. Take out the tissue sections and rinse them with phosphate buffered saline (PBS). Place the box containing the tissue sections and PBS on a shaker and rinse for 5 minutes each time for a total of 3 times.

[0066] 5. After the slices are rinsed, place them in an antigen repair solution at 100°C and heat them in a microwave oven. The antigen repair method used in the present invention is EDTA sodium solution (PH 9, purchased from Sevier Bio, item number G1203-250ML), the firepower is 30, and the time is 10 minutes. After the end, take out the antigen repair solution box and let it stand to cool naturally to room temperature. During this period, ensure that the antigen repair liquid surface completely covers the tissue.

[0067] 6. Place the slices in PBS (Phosphate buffered saline-Tween, abbreviated as PBST) supplemented with 5% polysorbate-20, and rinse in PBST on a shaker for 5 minutes each time, for a total of 3 washes.

[0068] 7. Place the tissue slices flat in a wet box. Use toilet paper to absorb the moisture on the surface of the slide. Then use a tissue pen to draw a circle 2mm around the edge of the tissue. Then add 3% hydrogen peroxide solution to the surface of the tissue and make sure the tissue is completely covered with hydrogen peroxide solution. After the end, cover the wet box and incubate it in a 37℃ biochemical incubator for 20 minutes.

[0069] 8. Remove the wet box and rinse the tissue sections with PBST for 5 minutes each time, for a total of 3 times.

[0070] 9. After rinsing, clean the surface of the tissue slice with toilet paper and place it in a wet box. Add about 50 μL of 3% goat serum (ready-to-use goat serum, purchased from Boster Biology, item number AR0009-12ML) to the surface of the tissue so that the tissue is completely covered with goat serum. Cover the wet box with a lid and place it in a 37°C biochemical incubator for 20 minutes.

[0071] 10. Dilute the primary antibody with PBS. The antibody dilution ratio is: EEF1E1, 1:200; ILVBL,

[0072] 1:200; SRPK1, 1:100; and NUDT5: 1:200. The prepared antibody dilutions were placed on ice for later use.

[0073] 11. Take out the wet box, use toilet paper to absorb the goat serum liquid on the surface of the tissue, and then drop about 50μL of the primary antibody dilution directly on the surface of the tissue. The antibody should completely cover the surface of the tissue. Then cover the wet box and incubate it at 4℃ overnight.

[0074] 12. After the primary antibody incubation for more than 12 hours, remove the wet box and rinse the tissue sections with PBST on a shaker. Rinse for 10 minutes each time, for a total of 3 times.

[0075] 13. After rinsing, use toilet paper to absorb the moisture on the surface of the slide, and then add secondary antibody (goat anti-rabbit secondary antibody for immunohistochemistry, purchased from Sevier Biotechnology, item number G1213-100UL) to the surface of the tissue. Ensure that the secondary antibody completely covers the tissue. Place the tissue sections in a wet box and incubate at room temperature for 30 minutes.

[0076] 14. After the secondary antibody incubation, remove the tissue sections and rinse them with PBST for 10 minutes each time, for a total of 3 rinses.

[0077] 15. Prepare DAB color development working solution (DAB color development kit, purchased from Sevier Biotechnology, item number G1212-200T), and add 50 μL of DAB color development solution to the tissue surface. Observe the cell coloring under a microscope. After sufficient coloring, immerse the slide in tap water to stop coloring.

[0078] 16. After all slides are stained, stain the nuclei with hematoxylin for 5 minutes. Then wash with tap water. Then put the slides into hydrochloric acid alcohol for differentiation for about 5 seconds, wash the slides with tap water again, and then immerse the slides in ammonia water to turn blue. Observe the coloring of the nuclei under a microscope. After the nuclei are stained, immerse the slides in tap water and rinse.

[0079] 17. Dehydrate the slides with gradient alcohol, the alcohol concentration is 75%, 85%, 95% in turn. Then immerse the slides in environmentally friendly dewaxing solution for 5 minutes to make them transparent. Finally, take out the slides and place them in a fume hood to dry.

[0080] 18. After the slides are dried, they are sealed with neutral resin. After the sealing is completed, the slides are dried and then taken under a microscope for observation and photography.

[0081] Tumor cell staining was scored using a semiquantitative five-category grading system: 0, no tumor cell staining; 1, 1-10% tumor cell staining; 2, 11-25% tumor cell staining; 3, 26-50% tumor cell staining; 4, 51-75% tumor cell staining; 5, >75% tumor cell staining. Staining intensity was scored using a semiquantitative four-category grading system: 0, no staining; 1, weak staining; 2, moderate staining; 3, strong staining. Tumor cell staining was scored by two independent experts who performed the staining under the same conditions at the same time. In the case of discordant scores, the sections were re-evaluated.

[0082] c) Immunohistochemical staining results

[0083] IHC confirmed that 4 proteins associated with hormone insensitivity (EEF1E1, ILVBL, SRPK1, and NUDT5 with available antibodies) were significantly increased in tissues of endometrial cancer patients with resistance to conservation therapy, further supporting the results that these proteins predict the response to progestin therapy. And correlation analysis showed that there was a positive correlation between the 4 proteins ( Figure 2 AC).

[0084] In summary, the present invention obtains 6 protein markers of resistance to conservation therapy of endometrial cancer through screening, and by detecting the expression levels of the 6 protein markers, the treatment response of patients with endometrial cancer can be predicted before the patients undergo conservation therapy, thereby effectively screening the drug-resistant population, and optimizing their treatment plans in a targeted manner, providing the patient with a benefit rate. The present invention constructs a detection product for predicting conservation resistance, which only requires collecting the patient's tumor tissue to perform the test according to the manual and evaluate whether the patient will develop drug resistance based on the staining results, which has the advantages of clinical accessibility and accuracy.

[0085] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0086] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A drug resistance marker for endometrial cancer conservation therapy, characterized in that: Including EEF1E1 protein, ILVBL protein, SRPK1 protein and NUDT5 protein; The drug for the conservation treatment of endometrial cancer is medroxyprogesterone acetate.

2. Application of drug resistance markers for conservation treatment of endometrial cancer, characterized in that: The reagent for detecting the drug resistance marker for conservation therapy of endometrial cancer as described in claim 1 is used to prepare a product for predicting drug resistance for conservation therapy of endometrial cancer, wherein the drug for conservation therapy of endometrial cancer is medroxyprogesterone acetate.

3. The use of the drug resistance marker for endometrial cancer conservation treatment according to claim 2, characterized in that: By quantitatively detecting the endometrial cancer conservation treatment resistance marker, the drug resistance status of patients with endometrial cancer conservation treatment can be judged.

4. The use of the drug resistance marker for endometrial cancer conservation treatment according to claim 2, characterized in that: The reagent for detecting the endometrial cancer conservation therapy resistance marker includes a substance that specifically binds to the EEF1E1 protein, the ILVBL protein, the SRPK1 protein and the NUDT5 protein.

5. The use of the drug resistance marker for conservation treatment of endometrial cancer according to claim 4, characterized in that: The substance is an antibody or an antibody fragment.

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