Application of THBS1 in preparation of pancreatic cancer diagnosis and treatment product

Abstract

The invention discloses application of THBS1 in preparation of pancreatic cancer diagnosis and treatment products. Wherein THBS1 in pancreatic cancer cells is high in expression, and the higher expression level of THBS1 is related to poor prognosis of pancreatic cancer patients; in addition, migration and invasion of pancreatic cancer cells can be inhibited by reducing THBS1 gene expression, and apoptosis is promoted at the same time; in addition, the THBS1 inhibitor can also be used for preparing a medicine for inhibiting a JAK2 / STAT3 signal channel (reducing phosphorylation of JAK2 and STAT3).

Description

Technical Field

[0001] The present application relates to the field of biomedical technology, and specifically to the use of a protein in preparing pancreatic cancer diagnosis and treatment products. Background Art

[0002] Pancreatic cancer (PC) is a powerful malignancy in the digestive tract, accounting for nearly 5% of cancer-related deaths worldwide. In 2022, approximately 511,000 new cases and 467,000 deaths will be attributed to PC worldwide.

[0003] The early diagnosis rate of PC is extremely low, and most patients have no obvious symptoms when the disease progresses to the late stage. PC is characterized by highly aggressive tumor cells, resulting in a 5-year survival rate of less than 10%. Known risk factors for PC include old age, smoking, diabetes, chronic pancreatitis, family history, obesity, excessive drinking, and genetic factors.

[0004] Treatments for PC include surgery, chemotherapy, targeted therapy, and immunotherapy strategies. Given the challenges of early diagnosis, most patients are diagnosed with PC at an advanced stage and are treated primarily with chemotherapy, although some patients may also consider immunotherapy or targeted drug therapy. Despite rapid progress in targeted therapy and immunotherapy, their clinical outcomes are unsatisfactory, and the effectiveness of treating PC remains limited. Therefore, there is an urgent need to study early diagnostic biomarkers and the molecular mechanisms of PC.

[0005] Thromboxane 1 (THBS1) is a secreted protein that functions in the tumor microenvironment. It has multiple functions, including inhibiting angiogenesis, regulating antitumor immunity, stimulating tumor cell migration, and regulating the activity of extracellular proteases and growth factors. Studies have shown that THBS1 contributes significantly to the development and progression of various cancers. Plasma THBS1 may be an unfavorable prognostic indicator for HER2-enriched breast cancer. Its increased expression is associated with poor outcome and immune response suppression in gastric cancer. THBS1 is involved in the PI3K / AKT signaling pathway and may play a role in oral squamous cell carcinoma. As a gene associated with high risk and prognosis, THBS1 may affect the initiation and progression of laryngeal cancer through the fatty acid metabolism pathway. These findings suggest that THBS1 may have a function in tumorigenesis. However, there are few studies on the function and mechanism of THBS1 in PC. Summary of the invention

[0006] This application clarifies the use of THBS1 in the preparation of pancreatic cancer diagnosis and treatment products.

[0007] In the study, the inventors used public databases to evaluate the expression level of THBS1 in PC and its association with prognosis, and conducted a series of in vitro and in vivo studies to examine the expression of THBS1 in PC cell lines and clarify its functional impact. These findings provide a theoretical and experimental basis for using THBS1 as a potential therapeutic target for PC treatment.

[0008] Therefore, this application provides the following practical technical solutions:

[0009] First, THBS1 is used in the preparation of products for diagnosis / screening of pancreatic cancer (pancreatic cancer can be screened using only THBS1 as a biomarker), wherein THBS1 is highly expressed in pancreatic cancer cells, and higher THBS1 expression levels are associated with poor prognosis in pancreatic cancer patients.

[0010] Optionally, the product for pancreatic cancer diagnosis / screening is a kit, a chip, a test strip or a computer program product. The computer program product here mainly means that when the user inputs the data of the THBS1 expression level of the sample to be tested, the computer program product can output the result of pancreatic cancer diagnosis / screening.

[0011] Secondly, THBS1 is used in the preparation of a product for treating pancreatic cancer, wherein pancreatic cancer cell migration and invasion are inhibited by reducing THBS1 gene expression, while apoptosis is promoted.

[0012] Optionally, the product for treating pancreatic cancer includes protein kinase inhibitors, RNAi and CRISPR systems.

[0013] Thirdly, the combination of THBS1 inhibitors and JAK2 / STAT3 signaling pathway activators is used in products for the treatment of pancreatic cancer. JAK2 / STAT3 signaling pathway activators generally promote the proliferation and metastasis of tumor cells. THBS1 inhibitors can be combined with JAK2 / STAT3 signaling pathway inhibitors for the development of therapeutic products for pancreatic cancer.

[0014] Fourthly, the use of THBS1 inhibitors in the preparation of drugs for inhibiting the JAK2 / STAT3 signaling pathway to reduce the phosphorylation of JAK2 and STAT3. Currently, the drugs available on the market that inhibit the JAK2 / STAT3 signaling pathway are mainly Ruxolitinib (Jakafi; Incyte / Novartis), Tofacitinib, Fedratinib, and Pacritinib. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1It is shown that THBS1 is overexpressed in pancreatic cancer (PC), and its expression is associated with the prognosis of PC patients;

[0016] Among them, (AB) According to the TCGA database, the expression of THBS1 in different cancer types (PAAD: pancreatic adenocarcinoma); (CD) According to the GTEx database, the abnormal expression of THBS1 mRNA and protein in PC; (E) According to the Kaplan-MeierPlotter database, the relationship between THBS1 expression and the prognosis of PC patients. (F) Compared with normal cells, the expression level of THBS1 in PC cells is higher. *p<0.05, ***p<0.001.

[0017] Figure 2 It shows that siTHBS1 inhibits the proliferation, migration and invasion of BXPC-3 cells, but promotes cell apoptosis;

[0018] Among them, (A) qRT-PCR results of THBS1 expression in BxPC-3 cells in different siTHBS1 treatment groups. (B) CCK8 test results of cell activity. (C) Results of cell apoptosis detected by cell analyzer. (D) qRT-PCR results of Bcl-2, Bax and Caspase3 mRNA expression. (E) Western blotting results of Bcl-2, Bax and Caspase3 protein expression. (FG) Transwell assay results of migration and invasion. (H) Western blotting results of JAK2 and STAT3 expression. *p<0.05, **p<0.01, ***p<0.001.

[0019] Figure 3 It shows that the JAK2 / STAT3 signaling pathway activator partially reversed the effect of siTHBS1 in BXPC-3 cells;

[0020] Among them, (A) CCK8 test results of cell activity under different concentrations of Colivelin treatment. (B) Western blotting results of JAK2 and STAT3 expression under different concentrations of Colivelin treatment. (C) CCK8 test results of cell activity after siTHBS1+Colivelin treatment. (D) Results of cell apoptosis detected by cell analyzer. (E) qRT-PCR results of Bcl-2, Bax and Caspase3 mRNA expression. (F) Western blotting results of Bcl-2, Bax and Caspase3 protein expression. (GH) Transwell assay results of migration and invasion. (I) Western blotting results of JAK2 and STAT3 expression after siTHBS1+Colivelin treatment. *p<0.05, **p<0.01, ***p<0.001.

[0021] Figure 4 To show that THBS1 knockdown inhibits the growth of pancreatic cancer tumors in mice;

[0022] Among them, (A) Fluorescence microscope used to observe the transfection efficiency of LV-shTHBS1. (B) qRT-PCR results of THBS1 expression after LV-shTHBS1 transfection. (C) Pictures of mice and tumor tissues in each group. (D) Weight of tumor tissue in each group. (E) Tumor growth curve (volume) in each group. (F) HE staining of tumor tissue. (GH) Tunel experiment of cell apoptosis in tumor tissue. (I) Western blotting results of E-cadherin and N-cadherin expression. (J) Western blotting results of JAK2 and STAT3 expression. *p<0.05, **p<0.01, ***p<0.001. DETAILED DESCRIPTION

[0023] Thromboxane 1 (THBS1) is a secreted protein that has been implicated in the progression of multiple cancers, but its specific contribution to pancreatic cancer (PC) has not been fully investigated. We investigated the association between THBS1 levels and PC prognosis. The cellular function of siTHBS1 was determined by in vitro functional experiments, including CCK8 assay for cell proliferation, Muse cell analyzer for apoptosis, and Transwell assay for invasion and migration. Colivelin was used in BXPC-3 cells for recovery experiments to investigate the mechanism by which THBS1 regulates the JAK2 / STAT3 pathway. In addition, subcutaneous tumors in nude mice were constructed using LV-shTHBS1 lentivirus to verify the function of THBS1 in vivo.

[0024] Results: Elevated THBS1 expression in PC was associated with poor prognosis. THBS1 was highly expressed in PC cells. siTHBS1 inhibited the growth, migration, and invasion of BXPC-3 cells, while promoting apoptosis. THBS1 inhibition also led to decreased phosphorylation of JAK2 and STAT3. The biological effects could be partially reversed by the JAK2 / STAT3 signaling pathway activator (Colivelin). In addition, shTHBS1 could inhibit the growth of implanted tumors in nude mice.

[0025] Conclusion: THBS1 knockdown inhibits cell proliferation, migration and invasion, and enhances cell apoptosis through the JAK2 / STAT3 signaling pathway.

[0026] The following is a detailed introduction to the research content and conclusion analysis related to this application plan.

[0027] Materials and methods:

[0028] 1. Expression and survival analysis of THBS1 in PC. Expression and survival analysis of THBS1 in PC was performed using GEPIA (http: / / gepia.cancer-pku.cn / ) and UALCAN (https: / / ualcan.path.uab.edu / ), two publicly accessible databases containing gene expression data of PC patients from TCGA, GTEx, and OMICS projects. THBS1 expression in PC tissue samples in the TCGA dataset was analyzed using the GEPIA and UALCAN online platforms. In addition, survival analysis of THBS1 was performed using the Kaplan-MeierPlotter database (http: / / kmplot.com).

[0029] 2. Cell culture, transfection, and colivelin treatment. Three human PC cell lines, including BXPC-3 (Cellverse, Shanghai, China, number iCell-h030), AsPC-1 (Cellverse, Shanghai, China, number iCell-h021), and PANC-1 (Procell, Wuhan, China, number CL-0184), and a human normal pancreatic ductal epithelial cell line (HPDE, BNCC, Beijing, China, number BNCC338221) were used in this study as a control group. The cell lines were cultured in their respective growth media—PRMI-1640 for BXPC-3, AsPC-1, and HPNE cells, and DMEM for PANC-1, each supplemented with 10% FBS and 1% penicillin-streptomycin—at 37°C in a 5% CO2 environment. The cell culture and transfection procedures followed the relevant guidelines and regulations.

[0030] 3. THBS1 expression was knocked down using siRNA. The interference sequence was designed by GenePharma (Shanghai, China). Blank siRNA was used as a negative control (siNC). The sequences of THBS1 interference are detailed in Table S1. Cells were seeded in 6-well plates at a density of 1×105 cells / mL and maintained in culture for 24 hours. Once the cells reached 60-80% confluence, Lipofectamine TM siTHBS1 or siNC was transfected into cells using the Invitrogen 2000 kit (Invitrogen, USA). The transfection effect was determined by qRT-PCR after 48 hours. siTHBS1-2 and siTHBS1-3 showed the most consistent and stable effects and were selected for further experiments.

[0031] 4. A recovery experiment was performed in BXPC-3 cells using STAT3 activator (Colivelin) to study the mechanism by which THBS1 regulates the JAK2 / STAT3 pathway.

[0032] 5. Cell Proliferation, Apoptosis, Invasion and Migration

[0033] The effect of siTHBS1 on cell proliferation was evaluated using a CCK8 kit (Sigma-Aldrich; Merck KGaA). BXPC-3 cells were seeded in a 96-well plate at a standard density of 2 × 103 cells per well. Subsequently, CCK-8 reagent was added to each well and maintained at 37 ° C for 2 hours. The absorbance of each well was measured at 450 nm using a microplate reader (BioTek, Winooski, USA).

[0034] use The effect of siTHBS1 on apoptosis of BXPC-3 cells was determined by cell analyzer (Milipore, USA). Cells were suspended in 100 μL of 1% BSA and mixed with an equal volume of apoptosis working solution. After 20 minutes of light scattering incubation, apoptotic cells were identified.

[0035] Cell migration and invasion were assessed using the Transwell assay. For cell motility analysis, the upper chamber was filled with 500 μL of MEM medium. After incubation at 37 °C for 24 h, the remaining cells in the upper chamber were removed. For invasion, the Transwell chamber was placed in a 24-well plate; the upper chamber was pre-coated with Matrigel and seeded with 2 × 105 cells in serum-free medium, while the lower chamber contained medium supplemented with 15% FBS. The cells were treated with a fixative solution of methanol and formaldehyde for 30 min. Subsequently, they were stained with a 0.1% crystal violet solution for 30 min. After staining, the cells were imaged and counted.

[0036] VI. Nude Mouse Tumor Formation Experiment

[0037] The shRNA sequences targeting THBS1 and lentiviral vectors (LV3) with green fluorescent protein (GFP) tags were purchased from GenePharma (Shanghai, China). The target sequences (shNC and shTHBS1-2 / 3) were ligated to the LV3 vector by T4 DNA ligase and then transduced into 5 × 105 BXPC-3 cells with a multiplicity of infection (MOI) of 100. Stable knockdown of shTHBS1 infected cell lines was selected with purulent oxime. After 96 h, the transfection efficiency was evaluated by fluorescence microscopy and qRT-PCR detection of THBS1 expression based on GFP expression.

[0038] Eighteen SPF BALB / c nude mice, aged 4-6 weeks, male, from SiPeiFu (Suzhou, China). These mice were randomly assigned to three groups of six, and a cell suspension containing 5×105 cells was subcutaneously injected in the axillary region, from shNC, shTHBS1-2 and shTHBS1-3 groups. The health and tumor growth of the mice were closely observed after inoculation. Tumor size was recorded every three days, and the volume was calculated using the formula V(mm3)=ab2 / 2. Tumor growth during the 5-week study was tracked using a growth curve. At the end of 5 weeks, mice were humanely killed, and subcutaneous tumors were removed to measure size and weight. All animal procedures were performed in accordance with the guidelines of the Animal Ethics Committee of Hainan Cancer Hospital. Finally, tumor tissue was sliced ​​and stained with hematoxylin-eosin (HE) and Tunel, both of which were from Beyotime (Shanghai, China).

[0039] 7. qRT-PCR

[0040] use Reagents were obtained from Thermo Fisher Scientific to extract RNA from cells and tissues, and their concentrations were determined using a Nanodrop-2000 spectrophotometer. RNA was then reverse transcribed into complementary DNA (cDNA) using the NovoScript 1st Strand cDNA Synthesis Kit. SYBR qPCR SuperMixplus (Novoprotein, Suzhou, China) was used for quantitative PCR, and GAPDH was used as a reference gene to normalize relative mRNA levels. The relative expression levels were calculated using the 2-ΔΔCt method. The primer sequences are detailed in Table S1.

[0041] 8. Western blot

[0042] Proteins were extracted from cell and tissue lysates and quantified using a BCA protein kit from Beyotime Biotechnology, Shanghai, China. Equal amounts of protein were separated on 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose membranes. The membranes were treated with primary antibodies overnight at 4°C to block nonspecific binding. The next day, the membranes were treated with horseradish peroxidase-conjugated secondary antibodies for 1 hour. Bands were detected using an enhanced chemiluminescence (ECL) detection system (YESEN, Shanghai, China) and analyzed using ImageJ software, with GAPDH as a loading control.

[0043] IX. Data Analysis

[0044] Data are presented as mean ± standard deviation (SD). Statistical comparisons were performed using two-way analysis of variance (ANOVA). Data analysis was performed using GraphPadPrism version 8.0, and P values ​​less than 0.05 were considered statistically significant.

[0045] Experimental results:

[0046] THBS1 is upregulated in PC. Figure 1 AB shows the results of GEPIA and UALCAN online tools showing the expression profile of THBS1 in various tumors, indicating that THBS1 is upregulated in PC. Analysis of the TCGA database showed that the transcript and protein levels of THBS1 were higher in PC tissues compared with adjacent non-cancerous tissues ( Figure 1 CD). In addition, we found that higher THBS1 expression levels were associated with poor prognosis in PC patients ( Figure 1 E). Examination of THBS1 expression in PC cells revealed that THBS1 was significantly upregulated in these cells compared to control HPNE cells ( Figure 1 F).

[0047] Biological effects of THBS1 knockdown in BXPC-3 cells.

[0048] siTHBS1-1, siTHBS1-2, siTHBS1-3, and siTHBS1-4 were transfected into BXPC-3 cells, and their efficiency was evaluated by qRT-PCR. The results showed that THBS1 expression was reduced after siTHBS1-2 and siTHBS1-3 transfection ( Figure 2 A). Therefore, these siRNAs were selected for further experiments.

[0049] CCK8 experiments showed that downregulation of THBS1 inhibited the proliferation of BXPC-3 cells ( Figure 2B). Using Muse flow cytometry, we observed that the apoptosis rate of BXPC-3 cells increased after siTHBS1 treatment ( Figure 2 C). qRT-PCR and Western blot experiments showed that THBS1 inhibition led to decreased Bcl-2 expression and increased Bax and Caspase3 expression ( Figure 2 DE). In addition, Transwell assay showed that BXPC-3 cells migrated after THBS1 knockdown ( Figure 2 F) and invasion ( Figure 2 G) decreased ability. Collectively, these findings indicate that siTHBS1 treatment inhibited the growth, motility, and invasiveness of BXPC-3 cells and enhanced apoptotic activity.

[0050] In addition, the effect of THBS1 knockdown on the JAK2 / STAT3 signaling pathway in BXPC-3 cells was examined. It was found that the phosphorylation levels of JAK2 and STAT3 were downregulated after siTHBS1 treatment ( Figure 2 H). These results suggest that siTHBS1 may inactivate the JAK2 / STAT3 signaling pathway in BXPC-3 cells.

[0051] JAK2 / STAT3 signaling pathway activators could partially reverse the effects of siTHBS1 in BXPC-3 cells.

[0052] Subsequently, to further confirm whether THBS1 knockdown inhibits BXPC-3 cell proliferation, migration, and invasion by inhibiting the JAK2 / STAT3 signaling pathway, a STAT3 activator (Colivelin) was used. Increased activity of BXPC-3 cells was observed under 1M, 2M, and 4M Colivelin treatment ( Figure 3 A). In addition, phosphorylation of JAK2 and STAT3 increased ( Figure 3 B). Therefore, 1 μM Colivelin was selected for further study.

[0053] Compared with BXPC-3 cells treated with siTHBS1, Colivelin-treated cells showed a significantly higher proliferation rate ( Figure 3 C) enhances cell apoptosis ( Figure 3 In addition, the expression levels of Bax and Caspase3 were downregulated, while Bcl-2 expression was upregulated in BXPC-3 cells treated with siTHBS1 after Colivelin treatment ( Figure 3 EF). Colivelin also partially alleviated the effect of siTHBS1 on cell migration ( Figure 3 G) and invasion ( Figure 3In addition, compared with BXPC-3 cells treated with siTHBS1, the phosphorylation of JAK2 and STAT3 increased after Colivelin treatment ( Figure 3 I). Collectively, these findings indicate that the JAK2 / STAT3 signaling pathway activator (Colivelin) can partially reverse the biological effects of siTHBS1 in BXPC-3 cells, suggesting that THBS1 may regulate PC via the JAK2 / STAT3 pathway.

[0054] shTHBS1 inhibits tumor formation in vivo:

[0055] To explore the biological role and potential mechanisms of THBS1 in vivo, we established a mouse model with subcutaneous tumor implantation. Figure 4 A) and qRT-PCR ( Figure 4 The results of B) indicated that LV-shTHBS1 was successfully transfected. After 5 weeks, the mice were sacrificed and the tumors were removed to measure the volume and weight. Compared with the LV-NC group, the tumor volume of the shTHBS1 group ( Figure 4 C) and weight ( Figure 4 D) decreased significantly, and the growth rate ( Figure 4 E) was also significantly reduced. H&E staining ( Figure 4 F) shows that after shTHBS1 treatment, tumor cells shrank and arranged loosely, pathological nuclear division decreased, and a small number of vacuoles and cell necrosis appeared in the tumor tissue. These results indicate that shTHBS1 can inhibit tumor growth in nude mouse models.

[0056] In addition, the Tunel experiment ( Figure 4 GH) showed that THBS1 knockdown promoted apoptosis. shTHBS1 treatment led to upregulation of E-cadherin and downregulation of N-cadherin in tumor tissues ( Figure 4 I). In addition, the expression of proteins related to the JAK2 / STAT3 pathway was evaluated. Consistent with the in vitro findings, shTHBS1 treatment reduced the phosphorylation of JAK2 and STAT3 ( Figure 4 J). These results suggest that shTHBS1 treatment inhibits PC cell migration and invasion in vivo by suppressing the JAK2 / STAT3 signaling pathway, while promoting apoptosis.

[0057] Here, bioinformatics analysis revealed that elevated THBS1 levels in PC were associated with adverse outcomes. Moreover, THBS1 expression was found to be higher in PC cells than in control HPNE cells. Furthermore, our findings showed that THBS1 knockdown inhibited proliferation, migration, and invasion of BXPC-3 cells via the JAK2 / STAT3 pathway in vitro and in vivo, but enhanced apoptosis. These results suggest that THBS1 could serve as a candidate biomarker for the diagnosis and treatment of PC.

[0058] Previous studies have shown that THBS1 has multiple functions, including protecting pancreatic β cells, regulating pancreatic stellate cells, and participating in PC progression. THBS1 is known to protect pancreatic β cells by maintaining the expression of midbrain astrocyte-derived neurotrophic factor. It also protects these β cells from lipotoxicity through the PERK-NRF2 pathway. In addition, microRNA let-7d targets THBS1 and inhibits the activation of human pancreatic stellate cells. Aberrant methylation of the THBS1 promoter region was found in the plasma of PC patients. Bioinformatics analysis showed that THBS1 is abnormally expressed in PC and may affect the prognosis of pancreatic ductal adenocarcinoma. In addition, circular RNA hsa_circ_0007919 promotes PC metastasis by regulating Sp1-mediated THBS1 transcription. However, there are few reports on the mechanism of THBS1 in PC. Our bioinformatics analysis showed that THBS1 levels were higher in PC tissues compared with adjacent normal tissues, and high expression was associated with a poor prognosis in PC. In addition, THBS1 was overexpressed in PC cells compared with control HPNE cells. Knockdown of THBS1 by siRNA inhibited the proliferation, migration, and invasion of BXPC-3 cells, while promoting apoptosis. In addition, shTHBS1 inhibited tumor growth implanted in nude mice, highlighting the potential of THBS1 as a therapeutic target for PC.

[0059] In our study, phosphorylation of JAK2 and STAT3 in cells and animal models was reduced after THBS1 knockdown treatment, indicating that THBS1 knockdown may inactivate the JAK2 / STAT3 pathway. Colivelin, a JAK2 / STAT3 signaling pathway activator, can partially reverse the biological effects of siTHBS1 in BXPC-3 cells, suggesting that THBS1 may regulate PC through the JAK2 / STAT3 pathway.

[0060] Therefore, THBS1 inhibitors can be used in combination with JAK2 / STAT3 signaling pathway inhibitors for the development of therapeutic products for pancreatic cancer. Accordingly, the combination of THBS1 inhibitors and JAK2 / STAT3 signaling pathway activators has the use of products for the treatment of pancreatic cancer.

[0061] In addition, the drugs currently available on the market that inhibit the JAK2 / STAT3 signaling pathway are mainly Ruxolitinib (Jakafi; Incyte / Novartis), Tofacitinib, Fedratinib, and Pacritinib. Based on the above experiments and analysis, it can also be confirmed that THBS1 inhibitors are useful in preparing drugs for inhibiting the JAK2 / STAT3 signaling pathway and reducing the phosphorylation of JAK2 and STAT3.

[0062] In summary, our study provides evidence that THBS1 is overexpressed in PC and is associated with adverse outcomes. The data from this study showed that silencing THBS1 inhibited the proliferation, migration, and invasiveness of BXPC-3 cells, while also enhancing apoptosis by inactivating the JAK2 / STAT3 pathway. In addition, shTHBS1 inhibited tumor formation in vivo. Compared with the currently reported studies on pancreatic cancer biomarkers, this application clarifies the use of THBS1 alone in the preparation of products for diagnosis, screening, and treatment of pancreatic cancer through rigorous and sufficient experiments and analysis, as well as the feasibility of developing therapeutic products for pancreatic cancer by combining THBS1 inhibitors with JAK2 / STAT3 signaling pathway inhibitors.

Claims

1. Use of THBS1 in the preparation of a product for diagnosis / screening of pancreatic cancer, wherein: THBS1 is highly expressed in pancreatic cancer cells, and higher THBS1 expression levels are associated with poor prognosis in pancreatic cancer patients.

2. The use according to claim 1, characterized in that The product for pancreatic cancer diagnosis / screening is a kit, a chip, a test paper or a computer program product.

3. Use of THBS1 in the preparation of a product for treating pancreatic cancer, wherein: It inhibits pancreatic cancer cell migration and invasion by reducing THBS1 gene expression and promotes apoptosis.

4. The use according to claim 3, characterized in that The products for treating pancreatic cancer include protein kinase inhibitors, RNAi and CRISPR systems.

5. Use of a combination of a THBS1 inhibitor and a JAK2 / STAT3 signaling pathway activator in a product for treating pancreatic cancer.

6. Use of THBS1 inhibitors in the preparation of drugs for inhibiting the JAK2 / STAT3 signaling pathway, thereby reducing the phosphorylation of JAK2 and STAT3.

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