Effects Of PDAC Stroma ESE3 Expression On Progression And Chemotherapy Resistance In PDAC

Background Pancreatic cancer is one of the worst malignan tumors,with rapid disease progression,high local invasion ability and distant metastasis tendency,and about 95% of them is Pancreatic ductal adenocarcinoma(PDAC)originating from ductal epithelium.Only about 20% of patients with PDAC can improve their overall survival(OS)by surgical resection,but the overall five-year survival rate remains low.It has become a major public health problem that threaten to human health.Due to the special anatomical location of the pancreas,it has brought great difficulties to the early detection and early diagnosis of PDAC.Therefore,it is of great practical significance to pay attention to the quality of life of the discovered PDAC patients.Gemcitabine(GEM)is considered to be a first-line chemotherapy for patients with advanced pancreatic cancer and can improve the quality of life of patients with pancreatic cancer.However,only a small number of patients with PDAC are sensitive to GEM,thus improving chemotherapy resistance and improving sensitivity to GEM in patients with PDAC has become a priority.The characteristic structure of PDAC is fibrosis.The most important component is Pancreatic Stellate Cells(PSC).PSC,activated and proliferated by changes in tumor microenvironment,aggravating fibrosis of pancreatic cancer,releasing a variety of cytokines and inflammatory factors promote tumor cell proliferation,metastasis.Therefore,plentiful stroma causes resistance to chemotherapy,ultimately leading to the progression of pancreatic cancer,affecting the OS in PDAC patients.Epithelial-specific Ets transcription factor(ESE3)is a nuclear transcription factor that regulates the expression of target genes through transcriptional activation or inhibition,thereby affecting a variety of cellular functions.Our previous study found that ESE3 protein expression was increased in pancreatic cancer stroma compared with that in normal pancreatic stroma.In this study,we will further clarify the effects of the increased ESE3 expression on biological behavior in pancreatic cancer and plus,we will analyze the relationship between ESE3 expression and survival,other clinical pathologic parameters.And its role in the resistance of pancreatic cancer to GEM,and initially explore its mechanism of action and reversed strategy.Method 1.ESE3 immunohistochemical staining was performed in the PDAC stroma tissues to analyze the expression level of ESE3 protein in PSC.2.The clinical and pathological data of PDAC patients were followed up to analyze the relationship between the expression of ESE3 and OS/DFS,clinicopathological parameters.3.Application of western blot and other experimental techniques were conducted to detect the expression changes in ESE3 after stimulation of IL-1? factor in human immortalized PSC and human primary PSC,and the localization of ESE3 in the nucleus and cytoplasm.We construct human PSC cell line stably overexpressing ESE3(PSCpCDH-ESE3),co-cultured with pancreatic cancer cell lines,and detect changes in invasion function of tumor cells;detect tumor cells apoptosis after treatment with GEM.4.A subcutaneous tumor formation model of PSCs and tumor cell lines was established.Combination of GEM and NF-?B inhibitor to observe tumor size,growth rate and degree of PDAC fibrosis.Result 1.Expression of ESE3 in PSC Pancreatic cancer tissue was staining ESE3 by IHC,the expression of ESE3 in PSC was increased compared with that normal PSC.2.ESE3 expression in PSC was associated with TNM stage(P=0.037),tumor size(P=0.041),pancreatic cancer biomarkers CA19-9(P=0.030),and CEA(P=0.010).Moreover,comparing with patients with low expression of ESE3,high expression of ESE3 group had lower overall survival(OS)(33.73 months vs.17.0 months)and lower recurrence-free survival(DFS)(33.70 months vs.14.0 months),and the difference was statistically significant.After the analysis of the Cox risk prediction model,the high expression of ESE3 in pancreatic cancer stroma was an independent risk factor for the survival and recurrence-free survival of pancreatic cancer.3.Detected by Western blot,the NF-?B p65 pathway was activated in human immortalized PSC and primary PSC after stimulation with human recombinant IL-1?,accompanied by increased expression of ESE3 and located in the nucleus of PSC.Coculture of PSC overexpressing ESE3 with pancreatic cancer cell lines showed that the invasive ability of tumor cells(P <0.05).After treatment with gemcitabine,the percentage of apoptosis in tumor cells co-cultured with PSC overexpressing ESE3 was decreased compared with that in conventional cultured tumor cells.The difference was statistically significant(P < 0.05),indicating that the increase in ESE3 protein expression in PSC plays an important role in the progress.4.After the subcutaneous tumor formation model of PSCs and tumor cell lines was established.Combine GEM with NF-?B inhibitor were administered.Compared with other the treatment group,tumor size was significantly reduced,growth rate was slowed,and fibrosis was low.Conclusion 1.Compared with normal pancreatic PSC,ESE3 is highly expressed in the pancreatic cancer PSC.2.High expression of ESE3 in PSC can be considered as an independent risk factor,affecting the survival and prognosis of patients with PDAC and playing an important role in tumor progression.3.Increased expression of ESE3 in PSC promotes its activation,resulting in PDAC progression,for example,increased the invasive ability and anti-apoptotic ability of pancreatic cancer cells,and plays a key role in chemoresistance of pancreatic cancer.4.NF-?B inhibitor combined with GEM may increase the sensitivity of pancreatic cancer to gemcitabine chemotherapy.