Effect Of S-adenosylmethionine(SAMe)on Proliferation And Migration Of Human Intrahepatic Cholangiocarcinoma Cell Line ICC-9810 And Its Mechanism

Objective:To investigate the effects of S-adenosylmethionine(SAMe)pretreatment on the growth and invasiveness of human intrahepatic cholangiocarcinoma ICC-9810 cell line;and cmyc and urokinase-type plasminogen activator(uPA)genes.Expression in ICC-9810 cells.Methods:Intrahepatic cholangiocarcinoma ICC-9810 cells(human intrahepatic cholangiocarcinoma cells)were cultured by in vitro cell culture technique,and the growth inhibition rate of cells was detected by MTT method;Different concentrations(0,2,4 mmol/L)of SAMe were added to the culture medium,and the apoptosis and cycle of each group were detected by flow cytometry.The cell invasion was detected by Transwell method.In addition,mRNA expression of the c-myc and uPA genes was detected by reverse transcription polymerase chain reaction(qPCR),and protein expression of the cmyc and uPA genes was detected by Western blotting(WB),MSP method was used to detect cmyc and uPA genes’s methylation in each group.Results:Treatment of human intrahepatic cholangiocarcinoma ICC-9810 cells with different concentrations of SAMe at 0.5～32mmol/L for 24,48 and 72 hours showed significant inhibition of cell growth and proliferation,with the increase of SAMe drug concentration and prolonged action time.The apoptosis rate of cancer cells also increased significantly(P<0.01);the number of cancer cells invaded gradually decreased,and the invasive rate also decreased(P<0.01).In addition,from the experimental results of cell cycle,it can be seen that with SAMe As the concentration increases,the G0/G1 phase can be inhibited and the S phase can be promoted.The results of qPCR showed that compared with the control group,the expression of cmyc mRNA increased significantly with the increase of SAMe concentration.The expression of uPA gene mRNA was the highest when SAMe concentration was 4mmol/L,and the expression level was the lowest when SAMe concentration was 2mmol/L;WB The results showed that the expression of cmyc protein decreased with the increase of drug concentration,and uPA protein was not detected,The status of uPA and cmyc genes’s methylation was not detected successfully.Conclusion:SAMe significantly inhibited the growth of human intrahepatic cholangiocarcinoma ICC-9810 cell line,significantly reduced the invasiveness of cancer cells,and positively correlated with the concentration of SAMe.SAMe can play a role in inhibiting cell growth and reducing invasiveness by participating in the regulation of the expression of cmyc gene and uPA gene,and can provide a new strategy for clinical research on drug therapy for intrahepatic cholangiocarcinoma.