Expression Of TCP1? In Human Pancreatic Ductal Adenocarcinoma And The Effects On The Biological Behavior Of Pancreatic Cancer Cell Line

BackgroundPancreatic cancer is a highly malignant tumor of digestive system and the sixth most common cause of cancer-related death globally.At present,the treatments of pancreatic cancer are mainly surgery,chemotherapy,radiotherapy,immunotherapy and other comprehensive treatments.However,the prognosis of Pancreatic cancer is still very poor,with a five-year survival rate of only 7%.Therefore,there is an urgent need to discover new effective treatments for pancreatic cancer.In order to overcome the shortcomings of traditional cancer treatments,more and more researchers are devoted to the study of molecular targeted therapy.The ideal molecular targeted therapy drugs can specifically select a cancer-causing target site in tumor cells to combine with it,thereby specifically inhibit the growth of cancer cells,and then lead to the death of tumor cells.Oncogene addiction provides a good biological basis for molecular targeted therapy and has become an important research field for anticancer drug development and treatment of cancer.We have identified in previous studies that quick loss of multiple short-lived proteins may be associated with acute apoptosis following acute inhibition of the addicted oncogenes in cancer cells,and T complex protein 1 subunit ?(TCT-1-alpha,abbreviated as TCP 1??CCT1)is one of these proteins.TCPla is a subunit that constitutes molecular chaperone complex TRiC(also known as TCP-1,CCT).Several studies have shown that TRiC subunits are overexpressed in a variety of tumor samples,including liver cancer,lung cancer,breast cancer,gastric cancer,colon cancer,etc.And it was found that multiple subunits play a crucial role in tumor development.However,little is known about the relevant studies on TCPla in pancreatic cancer.Therefore,this topic will focus on the role of TCP1? in the development of pancreatic cancer to provide theoretical and experimental basis for the pathogenesis and targeted therapy of pancreatic cancer.PurposesPancreatic cancer has insidious onset,rapid development and difficulty in early diagnosis and treatment and it has one of the worst prognoses of digestive system malignant tumor.Therefore,there is an urgent need to discover new and effective targeted therapies.This topic aims to elucidate the expression of TCP1? in pancreatic cancer tissue and its role in the development of pancreatic cancer.Methods(1)The expression of TCPla in pancreatic cancer tissue and normal pancreatic tissue was assessed by immunofluorescence.Then,PROGgeneV2 online analysis tool was used to analyze the relationship between TCP1? expression and patient prognosis.(2)Silencing the expression of TCP1? proteins in the MiaPaCa-2 and PANC-1 pancreatic cancer cell lines using small interfering RNA technology,then the effects of inhibition TCP1? expression on biological behaviors of pancreatic cancer cells such as proliferation,migration and invasion in vitro were detected by proliferation experiments(MTT),scratch experiments and transwell migration and invasion experiments.(3)After down-regulating the expression of TCP1? in pancreatic cancer cell line MiaPaCa-2 and PANC-1,we can observe whether it can induce acute apoptosis by inhibiting KRAS gene related signaling pathway alone or in combination.Results(1)Immunofluorescence results showed that TCP1? showed strong fluorescence expression in pancreatic cancer tissues and weak fluorescence expression in the corresponding adjacent normal pancreatic tissues.PROGgeneV2 online analysis showed that high TCP1? expression was associated with poor prognosis in pancreatic cancer patients.(2)The silencing effects of specific siRNA targeting TCP1? were assessed by western blot analysis in pancreatic cancer cell line MiaPaCa-2 and PANC-1.The results showed that the interference rates of the three siRNA-TCP1? on TCP1?expression in MiaPaCa-2 and PANC-1 cells were 51%,72%,69%and 59%,72%,44%,respectively.Among them,siRNA-TCP1?-2 had the best interference effect,which was statistically significant(P<0.05).(3)Silencing the expression of TCP1? significantly inhibited the proliferation,migration and invasion of pancreatic cancer cell lines miapaca-2 and panc-1 in vitro(P<0.05,P<0.05,P<0.05).(4)Silencing TCP1? expression significantly promoted acute apoptosis of pancreatic cancer cells.Different combinations of LY294002(50 M),PD98059(40 M)and CHX(10 g/ml)were used to treat miapaca-2 and panc-1 pancreatic cancer cell lines that had been silenced with transfected siRNA for TCP1? expression,and to detect apoptosis-specific cleavage of PARP after 6 hours of treatment.Western Blot results showed that inhibition of TCPla expression combined with MEK1/ERK1/2 inhibitor PD98059 and protein synthesis inhibitor CHX could significantly promote the acute apoptosis of pancreatic cancer cells.Conclusions(1)TCP1? is highly expressed in pancreatic cancer and is associated with poor prognosis.(2)Silencing the expression of TCP1? significantly inhibited the proliferation,migration and invasion of pancreatic cancer cells in vitro.(3)Silencing the expression of TCP1? can partially promote the occurrence of acute apoptosis of pancreatic cancer cells.