Mechanism Of FAT10 Reducing The Chemotherapy Sensitivity Of Pancreatic Cancer Cells To Gemcitabine Through EMT

Background:Pancreatic cancer is a highly malignant digestive system tumor,and its chemoresistance to gemcitabine is partially responsible for the poor prognosis of patients.Therefore,methods that can improve the sensitivity of pancreatic cancer to gemcitabine chemotherapy are of clinical significance.Moreover,identifying new therapeutic targets for the development of effective therapeutic methods and biomarkers for pancreatic cancer required exploration.The ubiquitin-like protein FAT10 is abnormally expressed in various malignancies and is associated with poor prognosis.Recently,studies have found that the high expression of FAT10 is associated with chemoresistance to cisplatin,carboplatin,and 5-fluorouracil.However,the specific role of FAT10 in pancreatic cancer and its relation to the sensitivity of gemcitabine chemotherapy remains unexplored.Objective:This study aimed to verify the expression of FAT10 in pancreatic cancer and its correlation with the prognosis of patients with pancreatic cancer.The effect of inhibiting the biological function of FAT10 in pancreatic cancer cells and the sensitivity of gemcitabine chemotherapy were determined through in vitro and in vivo experiments.Finally,the specific molecular biological mechanism of FAT10 affecting the chemosensitivity of pancreatic cancer cells to gemcitabine was explored.Methods:The difference in FAT10 gene expression in pancreatic cancer and normal tissues was analyzed using the GEPIA2 online tool,and the m RNA and protein expression levels of FAT10 in pancreatic cancer and paired paraneoplastic tissues were verified using immunohistochemistry,q RT-PCR and Western Blot.Further,the association between FAT10 protein expression and clinicopathological features and its impact on prognostic implications was analyzed.Subsequently,the effects of inhibiting FAT10 expression on the biological functions of pancreatic cancer cells and sensitivity to gemcitabine chemotherapy were analyzed using the Edu proliferation assay,AO/EB apoptosis assay,wound healing assay,and cell-counting kit-8(CCK-8)cell viability assay in nude mice.Finally,the mechanisms by which FAT10 regulates the sensitivity of pancreatic cancer cells to gemcitabine chemotherapy were explored using the ss GSEA algorithm,cell immunofluorescence,Western Blot,and CCK-8 cell viability assays.Results:The GEPIA2 online tool suggested that the expression level of the FAT10 gene in pancreatic cancer was significantly higher than that in normal pancreatic tissues using the TCGA and GTEx databases.Immunohistochemical staining showed that FAT10 was highly expressed in pancreatic cancer tissues,and high FAT10 expression was associated with late TNM staging and poor prognosis.q RT-PCR and western blot results also showed that FAT10 m RNA and protein were highly expressed in pancreatic cancer cells.Further in vitro experiments found that interfering with FAT10 expression inhibited the proliferation and migration of pancreatic cancer cells,promoted the apoptosis of cells,and increased the sensitivity of pancreatic cancer cells to gemcitabine therapy.In vivo experiments also suggested that the stable interference of FAT10 expression inhibited tumor growth and increased gemcitabine’s therapeutic effect.Subsequently,the ss GSEA algorithm revealed that the FAT10 gene was closely related to the epithelial-mesenchymal transition(EMT)pathway score in pancreatic cancer.Additionally,cell immunofluorescence experiments showed that the interference of FAT10 expression could lead to the up-regulation of E-cadherin protein expression and the down-regulation of Vimentin protein expression.Finally,reversion experiments showed that interfering with FAT10 expression increased the sensitivity of pancreatic cancer cells to gemcitabine treatment,but the addition of an activator of EMT(TGF-β)prevented this process.Conclusion:FAT10 is highly expressed in pancreatic cancer and is associated with its poor prognosis.Interfering with FAT10 expression inhibits pancreatic cancer cell proliferation and migration,promotes apoptosis,and increases the sensitivity of pancreatic cancer cells to gemcitabine treatment.Furthermore,FAT10 mediates the sensitivity of pancreatic cancer gemcitabine chemotherapy by regulating EMT.