Use of abhd17c in diagnosis, treatment and prognosis of kras mutant pancreatic ductal carcinoma

Abstract

The application provides application of ABHD17C in diagnosis, treatment and prognosis of KRAS mutant pancreatic ductal carcinoma. It is found that the expression level of ABHD17C protein in KRAS mutant pancreatic ductal carcinoma patients is significantly higher than that in KRAS wild type pancreatic ductal carcinoma patients and healthy controls, and is significantly negatively correlated with the expression level of ALOX15B. In addition, the overall survival and recurrence-free survival of KRAS mutant pancreatic ductal carcinoma patients with high expression of ABHD17C protein are significantly shorter than those of KRAS mutant pancreatic ductal carcinoma patients with low expression of ABHD17C protein, and the difference has statistical significance. Moreover, silencing or inhibiting the expression of ABHD17C protein can promote ferroptosis of KRAS mutant pancreatic ductal carcinoma cells, can significantly inhibit the occurrence and development of KRAS mutant pancreatic ductal carcinoma, can inhibit the growth of tumor, and can effectively prolong the overall survival. It is indicated that the ABHD17C protein can be used as a diagnostic and prognostic biomarker of KRAS mutant pancreatic ductal carcinoma, and can be used as a specific treatment target of KRAS mutant pancreatic ductal carcinoma.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of disease diagnosis and biological medicine, and particularly relates to application of ABHD17C in diagnosis, treatment and prognosis of KRAS mutant pancreatic ductal adenocarcinoma. BACKGROUND

[0002] Pancreatic ductal adenocarcinoma (PDAC) is an invasive malignant tumor with the characteristics of rapid progression, easy metastasis, poor prognosis and high mortality. At present, surgical resection and adjuvant chemotherapy are the main methods for the treatment of pancreatic ductal adenocarcinoma. Because most patients with pancreatic ductal adenocarcinoma have developed into clinical advanced cancer stage at the time of first diagnosis, only 15%-20% of patients can receive surgical treatment. However, patients who have undergone surgical resection of tumor tissue still face problems such as postoperative metastasis and local recurrence, and the 5-year survival rate of patients is less than 8%. Therefore, exploring the mechanism of the development of pancreatic ductal adenocarcinoma is helpful for the early diagnosis of pancreatic ductal adenocarcinoma and the development of targeted treatment strategies for pancreatic ductal adenocarcinoma.

[0003] KRAS, as a GTPase, can cycle between inactive GDP-bound and active GTP-bound forms, and is the most frequently mutated oncogene in cancer. In pancreatic ductal adenocarcinoma, 86%-90% of patients have KRAS gene mutations in the 12th codon in the tumor, including G12D (45%), G12V (35%), G12R (17%) and G12C (1%-2%). Currently, clinical trials have confirmed the anticancer effect of KRAS inhibitors such as MRTX1133, sotorasib and adagasib. It has been reported that KRAS mutation can activate multiple cancer signaling pathways, change intracellular metabolic pathways, cause an increase in reactive oxygen species (ROS), and accelerate iron-dependent lipid peroxidation processes. Lipid peroxides play an important role in ferroptosis, so in recent years, ferroptosis has become a hot strategy for cancer treatment, and it is of great significance to develop new targeted ferroptosis treatment for KRAS mutant pancreatic ductal adenocarcinoma (KRASmut-PDAC).

[0004] Since it was first reported in 2012, ferroptosis has attracted much attention as a unique non-apoptotic cell death form. Ferroptosis is caused by iron-dependent lipid peroxides and is regulated by multiple cellular metabolic pathways, including redox homeostasis, mitochondrial activity and lipid metabolism. Intracellular free iron or iron-containing enzymes react with polyunsaturated fatty acids to produce a large amount of membrane lipid peroxides. When these lipid peroxides cannot be metabolized through the glutathione (GSH)-glutathione peroxidase (GPX4) system, a large amount of lipid peroxides will accumulate on the cell membrane, eventually leading to cell membrane rupture and cell death.

[0005] The occurrence and development of cancer cells are related to abnormal inhibition of ferroptosis, but the specific mechanism still needs to be studied. Therefore, finding the related molecules of ABHD17C that regulate the sensitivity of cancer cells to ferroptosis in pancreatic ductal carcinoma and regulating the sensitivity of cancer cells to ferroptosis are of great importance for the early diagnosis of PDAC and the development of targeted therapy strategies for PDAC. SUMMARY

[0006] Based on this, the purpose of the present application is the application of ABHD17C in the diagnosis, treatment and prognosis of KRAS mutant pancreatic ductal carcinoma.

[0007] To achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows.

[0008] In a first aspect of the present application, the application of ABHD17C protein as a biomarker in the detection of KRAS mutant pancreatic ductal carcinoma is provided.

[0009] In a second aspect of the present application, the application of ABHD17C protein as a biomarker in the prognosis evaluation of KRAS mutant pancreatic ductal carcinoma is provided.

[0010] In a third aspect of the present application, the application of a reagent for detecting the content of ABHD17C protein in the preparation of a KRAS mutant pancreatic ductal carcinoma detection product is provided.

[0011] In a fourth aspect of the present application, the application of a reagent for detecting the content of ABHD17C protein in the preparation of a KRAS mutant pancreatic ductal carcinoma prognosis evaluation product is provided.

[0012] In some embodiments, the reagent includes a reagent for western blot detection, immunohistochemical detection.

[0013] In some embodiments, the reagent is a specific antibody against ABHD17C protein, preferably a monoclonal antibody.

[0014] In some embodiments, the product is a kit.

[0015] In a fifth aspect of the present application, the application of a reagent for silencing or inhibiting the expression of ABHD17C protein in the preparation of a drug for treating KRAS mutant pancreatic ductal carcinoma is provided.

[0016] In some embodiments, the application includes promoting the occurrence of ferroptosis in KRAS mutant pancreatic ductal carcinoma cells.

[0017] In some embodiments, the reagent for silencing or inhibiting the expression of ABHD17C protein includes shRNA, an ABHD17C inhibitor.

[0018] The present application finds that the expression level of ABHD17C protein in KRAS mutant pancreatic ductal carcinoma patients is significantly higher than that in KRAS wild type pancreatic ductal carcinoma patients and healthy controls, and is significantly negatively correlated with the expression level of ALOX15B. In addition, compared with KRAS mutant pancreatic ductal carcinoma patients with low expression of ABHD17C protein, KRAS mutant pancreatic ductal carcinoma patients with high expression of ABHD17C protein have significantly shorter overall survival and recurrence-free survival, and the difference is statistically significant. It shows that ABHD17C protein can be used as a diagnostic and prognostic biomarker for KRAS mutant pancreatic ductal carcinoma, which can more specifically and more sensitively predict the occurrence of pancreatic ductal carcinoma, detect pancreatic ductal carcinoma, predict disease progression, evaluate treatment effect, and guide drug use and prognosis evaluation.

[0019] In addition, silencing or inhibiting the expression of ABHD17C protein can promote ferroptosis of KRAS mutant pancreatic ductal carcinoma cells, significantly inhibit the occurrence and development of KRAS mutant pancreatic ductal carcinoma, inhibit tumor growth, and effectively prolong overall survival. Therefore, ABHD17C protein can be used as a therapeutic target for KRAS mutant pancreatic ductal carcinoma. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 For the results of label-free quantitative proteomics analysis.

[0021] Figure 2 ABHD17C is highly expressed in KRAS mut -PDAC cells.

[0022] Figure 3 For the results of immunoprecipitation experiment.

[0023] Figure 4 ABHD17C is negatively correlated with the expression level of ALOX15B.

[0024] Figure 5 ABHD17C promotes down-regulation of ALOX15B expression by mediating depalmitoylation of ALOX15B.

[0025] Figure 6 KRAS mut Mitochondria in PDAC cells.

[0026] Figure 7 Immunohistochemical staining of PDAC xenograft tumor.

[0027] Figure 8 Standardized BLI signal of mouse PDAC.

[0028] Figure 9Representative BLI images for mice and Kaplan-Meier survival curves.

[0029] Figure 10 Representative images of ABHD17C expression in different PDAC tissues.

[0030] Figure 11 ABHD17C high expression and low expression KRAS mut Kaplan-Meier survival curves of PDAC survival curves. DETAILED DESCRIPTION

[0031] In order to facilitate the understanding of the present application, the present application will be described more fully below with reference to the embodiments. The preferred embodiments of the present application are given below. However, the present application can be realized in many different forms and is not limited to the embodiments described herein. The purpose of providing these embodiments is to make the disclosure of the present application more thorough and comprehensive. It should be understood that the experimental methods in the following examples are not specified, which are usually carried out according to the conventional conditions, or according to the conditions recommended by the manufacturer. The various reagents used in the examples are commercially available products.

[0032] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the application herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0033] The following will be described in conjunction with specific examples.

[0034] KRAS mutant pancreatic ductal carcinoma: KRAS mut -PDAC; KRAS wild-type pancreatic ductal carcinoma: KRAS wt -PDAC.

[0035] In the following examples, overexpression or knockout of genes is entrusted to Shandong Weizhen Biotechnology Co., Ltd. Among them, the ABHD17C-sh1 sequence for ABHD17C gene is: CATCAACTGTAACCAT ATAAA (SEQ ID NO. 1); the ABHD17C-sh2 sequence is: GCGTGAGTCCCGAGAA CATTA (SEQ ID NO. 2).

[0036] Example 1

[0037] Application of ABHD17C in the diagnosis of KRAS mutant pancreatic ductal carcinoma.

[0038] KRAS mut Tumor cells for culturing KRAS mut -PDAC organoids were obtained from Sun Yat-Sen University Cancer Center with the clinical ethics number B2022-639-01.

[0039] Fresh tumor tissues resected by surgery were immediately digested with 2 mg / ml collagenase at 37°C for 12 hours. The dissociated cells were embedded in growth factor-reduced Matrigel, followed by Matrigel solidification in 48-well plates and immediate administration of PDAC organoid medium. The medium was replaced every 2-3 days. Cell viability was detected using the CellTiter-Glo 3D kit (purchased from Promega).

[0040] Knocking out or overexpressing genes of organoids: collagenase digestion of organoids resulted in a single-cell suspension. The corresponding lentivirus (constructed by Shandong Weizhen Biotechnology Co., Ltd. on commission) and polybrene were added to the suspension, centrifuged at 300g for 1 hour at room temperature, and then incubated at 37°C for 3 hours. Cells were plated in Matrigel and continued to be cultured with PDAC organoids.

[0041] To explore the role of proteins in the development of KRAS mut -PDAC, we performed untagged quantitative proteomic analysis on 3 KRAS mutant PDAC tissues (one KRAS G12V -PDAC tissue and two KRAS G12D -PDAC tissues) and 3 KRAS wild-type (KRAS wt )-PDAC tissues to analyze the abundance of proteins in these PDAC tissues. The results are shown in Figure 1 a. The results showed that compared with KRAS wt -PDAC, a total of 82 proteins were abnormally expressed in KRAS mut -PDAC, of which 40 proteins were down-regulated and 42 proteins were up-regulated (P<0.01, log2 fold change>5).

[0042] Subsequently, we explored the effect of the above-mentioned 40 down-regulated proteins on the growth rate of organoids formed by KRAS mut -PDAC and KRAS wt -PDAC. The results are shown in Figure 1 b. Overexpression of ALOX15B had the most obvious inhibitory effect on the growth of 3 KRAS mut -PDAC organoids. Therefore, down-regulation of ALOX15B expression may be related to the development of KRAS mut -PDAC.

[0043] Further, KRAS wt -PDAC and KRAS mut -PDAC cells. KRAS wt -PDAC cells and KRAS mut -PDAC cells were immunoprecipitated and then subjected to mass spectrometry detection and analysis.

[0044] KRAS wt -PDAC cells and KRAS mut -PDAC cells were lysed with lysis buffer, placed on ice for 30 minutes, then centrifuged at 12000 rpm at 4°C for 10 minutes, and the supernatant was incubated with G protein agarose beads coupled with the corresponding antibody at 4°C overnight. The immunoprecipitate was washed 5-6 times and used for mass spectrometry analysis.

[0045] The results of mass spectrometry detection and analysis are shown in Figure 2 . Figure 2 a is that compared with KRAS wt -PDAC cells, the expression of 7 proteins binding to ALOX15B in KRAS mut -PDAC cells was significantly increased.

[0046] Figure 2 b is that western blot analysis showed that when the expression of the above 7 genes was knocked down, only when ABHD17C or DCAF10 was knocked down, the expression and half-life of ALOX15B could be significantly down-regulated.

[0047] Subsequent immunoprecipitation experiments found that Figure 3 ABHD17C and DCAF10 did not have a direct binding relationship, and knocking out ABHD17C significantly inhibited the binding between DCAF10 and ALOX15B, but knocking out DCAF10 did not affect the binding of ALOX15B and ABHD17C, indicating that the down-regulation of ALOX15B expression mediated by ABHD17C was achieved by regulating the binding between ALOX15B and DCAF10. In addition, knocking out DCAF10 significantly inhibited the binding of ALOX15B-DDB1 and ALOX15B-CUL4A, indicating that the CUL4 / DDB1 / DCAF10 ubiquitin ligase complex combined with ALOX15B and regulated the expression of ALOX15B. That is, ABHD17C can regulate the content of ALOX15B by affecting the ubiquitination and degradation of ALOX15B, thereby affecting ferroptosis leading to the occurrence and development of PDAC, suggesting that ABHD17C can be applied to the diagnosis of KRAS mut -PDAC.

[0048] Furthermore, 95 KRAS samples were collected. mut -PDAC patient tissue and 12 KRAS cases wt - PDAC patient tissues were fixed in formalin, embedded in paraffin, and sectioned. Immunohistochemical staining (IHC) was performed to detect the expression of ABHD17C and ALOX15B in each tissue. ABHD17C and ALOX15B scores were calculated based on the results, and the correlation between their expression levels was analyzed. Two independent pathologists reviewed and scored the immunostaining degree of formalin-fixed and paraffin-embedded sections based on the pathological characteristics of the tissue sections and patient data. The score was determined by a combination of the proportion of positively stained tumor cells and the staining intensity. The scores given by the two independent pathologists were combined into an average score for further comparative evaluation. Tumor cell proportion score: 0, no positive tumor cells; 1, positive cells <10%; 2, tumor cells positive 10-35%; 3, tumor cells positive 35-75%; 4, tumor cells positive 75%. Staining intensity was graded according to the following standards: 1, no staining; 2, weak staining (pale yellow); 3, moderate staining (yellowish brown); 4, strong staining (brown). The staining index (SI) was calculated as the product of the staining intensity score and the proportion of positive tumor cells. Using an assessment method, we evaluated protein expression in pancreatic ductal carcinoma tissue by measuring SI, with scores of 0, 2, 3, 4, 6, 8, 9, 12, and 16. SI ≥ 8 indicated high expression, and SI < 8 indicated low expression.

[0049] The results are as follows Figure 4 As shown, Figure 4 In the middle, 'a' represents two representative KRAS cases. mut -PDAC IHC staining image, size bar, 50 μm. Figure 4 b is KRAS mut -PDAC (n=95) and KRAS wt Data analysis plot of IHC results for -PDAC (n=12). The results show that ABHD17C in KRAS mut Increased expression in PDAC, and a negative correlation with ALOX15B expression levels, further confirms that ABHD17C can be applied to KRAS. mut -Diagnosis of PDAC.

[0050] Example 2

[0051] High expression of KRAS by ALOX15B wt Cells or ALOX15B with low KRAS expression wtThe cells were treated by overexpression or knockout of ABHD17C, and then the total expression level and palmitoylation level of ALOX15B were analyzed by ABE / IB experiment. The immunoprecipitates obtained in the immunoprecipitation experiment were separated by SDS-PAGE to separate the proteins, and then the proteins were transferred to a nitrocellulose membrane or a PVDF membrane, incubated with the corresponding specific antibodies to bind the target proteins, and then incubated with horseradish peroxidase or alkaline phosphatase-labeled secondary antibodies, and finally developed to obtain the target protein band.

[0052] Figure 5 In a, the total expression level and palmitoylation level of ALOX15B after the specific cells were treated by overexpression or knockout of ABHD17C, the results showed that in KRAS wt In PDAC, overexpression of depalmitoylase ABHD17C would cause a significant decrease in the total expression level and palmitoylation level of ALOX15B; while in KRAS mut In PDAC, silencing ABHD17C or inhibiting ABHD17C activity with ABD975 would cause an increase in the total expression level and palmitoylation level of ALOX15B.

[0053] In KRAS wt cells with high expression of ALOX15B or KRAS wt cells were treated by overexpression or knockout of ABHD17C, and then ALOX15B was subjected to IF staining. As shown in Figure 5 In b, it can be seen that in KRAS wt In PDAC, overexpression of ABHD17C would cause a significant decrease in the membrane localization of ALOX15B; while in KRAS mut In PDAC, silencing ABHD17C or inhibiting ABHD17C activity with ABD975 would cause an increase in the membrane localization of ALOX15B, suggesting that ABHD17C regulates the down-regulation of its expression by mediating the depalmitoylation of ALOX15B.

[0054] Further, after the KRAS mut ABHD17C gene in PDAC was knocked out using shRNA, and then the cells were treated with 5 μM Erastin or 10 μM RSL3, and the transmission electron micrographs of the mitochondrial crista in the cells were obtained. The results showed that Figure 6) showed typical ferroptosis morphological features, especially the disappearance of mitochondrial cristae, confirming that ABHD17C can inhibit ferroptosis of PDAC. Previous studies have reported that ALOX15B can convert arachidonic acid into 15-HpETE to induce ferroptosis, suggesting that ABHD17C plays an important role in the ferroptosis of pancreatic ductal carcinoma cells by regulating the expression level of ALOX15B.

[0055] Example 3

[0056] A PDAC xenograft tumor mouse model was constructed as follows: 4-6 week old NOD-SCID IL-2rγ- / - (NOG) mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were divided into 6 groups, 6 mice in each group, and treated as follows: each group of mice was injected with 5x10 5 sh-V KRAS mut -PDAC cells, ABHD17C-sh#1 KRAS mut -PDAC cells and ABHD17C-sh#2 KRAS mut -PDAC cells. Doxycycline (DOX, which can regulate gene expression in the Tet-on system, gene knockout does not occur without tetracycline, and only when tetracycline is given will it lead to the knockout of specific genes in specific locations) was dissolved in mouse drinking water at a concentration of 0.5-8 mg / ml and administered continuously for 12 weeks, then MLS000545091 (50 mg / kg) was administered by intraperitoneal injection. The mice were injected with D-luciferin (75 mg / kg) every week, and the tumor growth condition was observed and evaluated by bioluminescence imaging technology (IVIS Spectrum In Vivo Imager).

[0057] After 12 weeks, the pancreatic tissues of the mice in each group were taken, and the expression levels of 3 ALOX15B metabolites, 2 lipid peroxide products and caspase-3 were detected using IHC staining.

[0058] Figure 7 a is a representative IHC staining image obtained after corresponding treatment of PDAC xenograft tumor, size bar, 50 μm. b is the expression level of 3 metabolites of ALOX15B, 15-HETE, 13-HODE and 15-HPEP. The results show that compared with the control group tumor, the KRAS mut- ALOX15B metabolites (e.g., 15-HETE, 13-HODE and 15-HPEP) and 2 lipid peroxidation products (4-HNE and MDA) were significantly increased in tumors formed by PDAC cells, and caspase-3 levels were unchanged, confirming that ABHD17C mediates KRAS mut - sensitivity to ferroptosis in PDAC cells, thereby mediating KRAS mut - progression of PDAC.

[0059] Example 4

[0060] A PDAC orthotopic tumor mouse model was constructed, and the method, grouping and treatment of each group were the same as in Example 3.

[0061] After 12 weeks, the relative changes in BLI of mice in each group were detected.

[0062] Figure 8 The normalized BLI signal of orthotopically inoculated mice with specified PDAC cells was treated with DOX and ALOX15B inhibitor MLS000545091 (50 mg / kg), n = 6 / group. When the BLI signal reached 3.0 x 10 6 p / s / cm 2 / sr, the mice were injected intraperitoneally with DOX, which can significantly inhibit the expression of ABHD17C and induce the expression of ALOX15B. The results showed that injection of DOX can significantly inhibit the growth of orthotopic KRAS mut - PDAC, suggesting that ABHD17C can promote the KRAS mut - PDAC development.

[0063] Figure 9 Figure 8 is a representative BLI image of orthotopically inoculated mice with specified PDAC cells, scale bar, 50 μm. Figure 9 Figure 9 is a Kaplan-Meier survival curve of mice in each group. These results demonstrate that ABHD17C promotes the KRAS mut - PDAC development, while silencing ABHD17C in KRAS mut - PDAC cells can significantly inhibit the occurrence and development of PDAC and effectively prolong the overall survival of mice, indicating that ABHD17C is an important target for inhibiting KRAS mutant pancreatic ductal carcinoma.

[0064] Example 5

[0065] In this example, the expression level of ABHD17C in different tissue samples was verified. Eight cases of normal pancreatic tissue, 12 KRAS wt - PDAC tissues and 95 KRASmut -PDAC tissues, formalin-fixed patient tissues were then paraffin-embedded, sectioned, and finally subjected to immunohistochemical staining to detect the expression level of ABHD17C in each group of tissues.

[0066] Figure 10 ABHD17C in normal pancreatic tissues (n=8), KRAS wt -PDAC tissues (n=12) and KRAS mut -PDAC tissues (n=95) (left) and quantification (right). Scale bar, 50 pm. The results show that the expression level of ABHD17C in KRAS wt -PDAC tissues is significantly higher than that in normal pancreatic tissues and KRAS mut -PDAC tissues. mut -PDAC.

[0067] Example 6

[0068] Ninety-five cases of KRAS mut -PDAC tissues were collected, and the expression level of ABHD17C in each tissue was detected by immunohistochemical staining. According to the detection results, KRAS mut -PDAC patients were divided into KRAS mut -PDAC patients with high expression of ABHD17C and KRAS mut -PDAC patients with low expression of ABHD17C.

[0069] According to the above grouping standard, KRAS mut -PDAC patients with high expression of ABHD17C (high expression 17C) were 64 cases, and KRAS mut -PDAC patients with low expression of ABHD17C (low expression 17C) were 31 cases. Kaplan-Meier analysis of survival curves and recurrence-free survival curves of the two groups of patients was performed.

[0070] Figure 11 KRAS mut -PDAC patients with low expression of ABHD17C and KRAS mut -PDAC patients with high expression of ABHD17C (n=95; P<0.001, log-rank test). The results show that the survival curve and the recurrence-free survival curve of KRAS mut -PDAC patients with high expression of ABHD17C are significantly better than those of KRAS mutPatients with PDAC had significantly shorter overall survival and relapse-free survival, with statistically significant differences. These results indicate that ABHD17C can be used as a KRAS... mut - ABHD17C expression level is a prognostic biomarker for PDAC patients and can be used to assess KRAS. mut - Prognosis of PDAC patients.

[0071] Furthermore, we examined 54 cases of KRAS. mut -PDAC tissue analysis was performed to verify that ABHD17C expression levels can be used to assess KRAS. mut - Prognosis of PDAC patients.

[0072] Immunohistochemical staining was used to detect the expression level of ABHD17C in various tissues. Based on the results, KRAS... mut -PDAC is divided into KRAS with high expression of ABHD17C. mut - PDAC patients and KRAS with low ABHD17C expression mut For PDAC patients, the criteria for determining high and low expression are the same as above.

[0073] The results are as follows Figure 11 As shown, consistent with the above results, KRAS with low ABHD17C expression... mut Compared to PDAC patients, KRAS with high ABHD17C expression mut Patients with PDAC had significantly shorter overall survival and relapse-free survival, with statistically significant differences. This further validates that ABHD17C can be used as a KRAS (Korean Respiratory Syndrome Regulator). mut - Biomarkers for prognostic assessment of PDAC patients.

[0074] Based on the above experimental data, ABHD17C protein can serve as a diagnostic and prognostic biomarker for KRAS-mutant pancreatic ductal carcinoma, and as a target marker for KRAS-mutant pancreatic ductal carcinoma.

[0075] To achieve precise treatment of PDAC.

[0076] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0077] The above embodiments only express several implementation manners of the present application, and the description is more specific and detailed, but it should not be understood as a limitation on the patent scope of the present application. It should be noted that for ordinary skilled persons in the art, without departing from the concept of the present application, several modifications and improvements can be made, which are all within the protection scope of the present application. Therefore, the protection scope of the patent of the present application should be subject to the appended claims.

Claims

1. Application of reagents for detecting ABHD17C and ALOX15B protein levels in the preparation of KRAS-mutant pancreatic ductal carcinoma detection products.

2. Application of reagents for detecting ABHD17C and ALOX15B protein levels in the preparation of prognostic assessment products for KRAS-mutant pancreatic ductal carcinoma.

3. The application as described in claim 1 or 2, characterized in that, The reagents include those used for Western blot detection and immunohistochemical detection.

4. The application as described in claim 3, characterized in that, The reagent used to detect the ABHD17C protein content is a specific antibody against the ABHD17C protein.

5. The application as described in claim 4, characterized in that, The specific antibody is a monoclonal antibody.

6. The application as described in any one of claims 1 to 5, characterized in that, The product in question is a reagent kit.

7. The use of a reagent that silences or inhibits ABHD17C protein expression as the sole active ingredient in the preparation of a drug for treating KRAS-mutant pancreatic ductal carcinoma, characterized in that, The reagent is shRNA, and the nucleotide sequence of the shRNA is shown in SEQ ID NO. 1 or SEQ ID NO.

2.

8. The application as described in claim 7, characterized in that, The applications include promoting ferroptosis in KRAS-mutant pancreatic ductal carcinoma cells.

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