The Role And Related Mechanism Of TRPV6 In Proliferation And Apoptosis Of Pancreatic Cancer

ObjectivePancreatic cancer is the highly frequently occuring malignant disease with the poor prognosis,and more than 90% patients will come to death in 1 year and 5-year survival rate is less than 5%,and prognosis cound not be elvated in the past 30 years.TRPV6 is the highly selected calcium channel protein,which could participate in the development of various tumors.The aim of study is to elucidate the influence of TRPV6 on the pancreatic cancer,and role and the related mechanism of TRPV6 in the proliferation and apoptosis of pancreatic cancerMethodsFrom Jan.2006 to Jan.2014,there were 76 pairs of sample of pancreatic ductal adenocarcinoma tissue and adjacent noncancerous tissue plus 6 pairs of pancreatic cancer samples and its counterpart adjacent noncancerous tissue to detect.(1)IHC and WB were used to detect the expression of TRPV6 to investigate the raltionship between cliniopathological parameter and prognosis.(2)Cell culture,AsPC-1,Bx PC-3,Capan-2,sw1990,PANC-1 were cultured.(3)WB was used to detect the protein expression of TRPV6 in the various pancreatic cancer cells.(4)Cell Counting Kit-8(CCK8)was used to detect the proliferative inhibitory effect of TRPV6-Si in pancreatic cancer cells(Capan-2,sw1990).(5)Flow cytometry(FCM)was to detect the pancreatic cancer cell cycle and the apoptosis after use of TRPV6-SiRNA.(6)Transwell was to detect the migration and invasion of pancreatic cancer cell(7)Western blot was to detect the target protein expression of its related proliferative and apoptotic signaling pathway.(8)Combination of TRPV6-SiRNA and 3 differentchemical drug to detect the sensitivity of drug.Results(1)IHC: TRPV6 was highly detected in malignant tissues.Positive expression of TRPV6 was related to the tumor size and TNM stage.(2)Kaplan-Meier analysis showed that a possitive expression of TRPV6 was associated with the poor prognosis.Multivariate analysis showed vessel invasion and TRPV6 were the independent risk factors correlated with the prognosis.WB: the protein expression of the pancreatic cancer is significantly higher than the counterpart normal tissue in 6 cases.(3)CCK8: the growth inhibitory effect is definite by TRPV6-SiRNA(4)FCM: the rate of apoptosis effect is promoted by TRPV6-SiRNA;G1 was arrested in the cell cycle.(5)TRPV6-SiRNA inhibited the migration and invasion of pancreatic cancer cell(6)TRPV6-SiRNA decreased the level of Numb?the antiapoptotic protein bcl-2?MMP9,and increased the level of the proapoptotic protein Bax?E-cadherin(7)Si-TRPV6 could increase the sensitity of oxalipalitin.ConclusionThe tumor size and TNM stage were associated with the expression of the TRPV6 in pancreatic cancer,the patients could get the better prognosis with lower TRPV6 expression.The TRPV6-SiRNA may inhibit the proliferation of the pancreatic cancer cell,induce apoptosis,arrest G1 and inhibit the invasion and migration.TRPV6-SiRNA could increase the sensitity of oxalipalitin.TRPV6 can regulate the biological behavior of pancreatic cancer cell via PCNA,Bcl-2,BAX,E-cadherin,MMP9.