The Research About SiRNA Targeting Notch1 Affected Biological Behavior Of Intrahepatic Cholangiocarcinoma Cells

Objective : Investigated the expression of Notch1 in intrahepatic cholangiocarcinoma and adjacent tissues;HCCC-9810,RBE and HIBEC cells.Small interfering RNA(siRNA)Targeting Notch1 were transfected into HCCC-9810 cells In order to study the effects on the proliferation,expression,apoptosis and invasion.At the same time,we explore the mechanism of it.Method:(1)Normal and ICC tumor tissue(confirmed by clinical pathology)samples were collected for analysis.Human LncRNAAngilent 4*180k Chip was used to screen LncRNA and mRNA co-expression analysis in order to study Notch gene’s expression;The protein levels of notch1 in intrahepatic cholangiocarcinoma cell line HCCC-9810 RBE and human intrahepatic biliary epithelial cell line HIBEC were evaluated by western blot analysis.(2).In order to estimate the effects of small interfering RNA(siRNA)targeting notch1 on the proliferation,expression,apoptosis and invasion abilities of intahepatic choangiocarcinoma,siRNA targeting notch1 and negative control siRNA were transfected intahepatic choangiocarcinoma cell line HCCC-9810.The mRNA levels of notch1 in the transfected cells were estimated by reverse transcription-quantitative polymerase chain reaction(RT-qPCR),and the protein levels of notch1 in these cells were evaluated by western blot analysis.The growth,apoptosis and invasion abilities of the transfected cells were measured by Cell Counting Kit-8(CCK-8),Annexin V-FITC Apoptosis Detection Kit)and transwell chamber assay,respectively.(3)The link between notch singal pathway and EMT was measured by western-blot.Result: Notch1 protein expression in intrahepatic cholangiocarcinoma cell lines was significantly higher than non-cancerous tissue.intrahepatic cholangiocarcinoma cell lines HCCC-9810 and RBE were significantly higher than normal biliary epithelial cell line HIBEC.Notch1 siRNA can downregulate the transcription of Notch1 mRNA and the expression of Notch1 protein.Downregulate Notch1 protein expression in intrahepatic cholangiocarcinoma cell line HCCC-9810 can significantly affect their proliferation,apoptosis,invasion and metastasis(p﹤0.05).Depth study found lower levels of Notch1 protein expression in intrahepatic cholangiocarcinoma cell line HCCC-9810 impacts on the invasion and metastasis may be associated with EMT.Conclusion:Notch1 proteins in intrahepatic cholangiocarcinoma are abnormally high expression.Interference silencing Notch1 protein expression in intrahepatic cholangiocarcinoma cell line HCCC-9810 cells can inhibit their proliferation,invasion and metastasis,while promoting their apoptosis.Notch signal pathway promotes cell invasion and metastasis may be associated with EMT.This suggests that Notch signaling pathway has an important influence on the biological behavior of intrahepatic cholangiocarcinoma and may become a new therapeutic target.