Effect Of RNAi-mediated Gene Silencing Of Mcl-1 On Apoptosis Of Human Pancreatic Cancer Cell Line PANC-1

Objective By RNAi technology Mcl-1 in Human pancreatic cancer cell line PANC-1 was specifically knocked down with siRNA, then detect and analyze the impact on proliferation and apoptosis of PANC-1 cells in vitro. From this research we hope to provide the experimental foundation for gene treatment of the pancreatic cancer.Methods 1 The lipofectamine ? 2000 and FAM-NC-siRNA were mixed in different proportions then transfected into PANC-1 cells. Transfection efficiency was detected by flow cytometry, and then select the best transfection conditions. 2 The inhibition of Mcl-1 expression was measured by reverse transcription PCR (RT-PCR) and Westernblot after transfecting siRNA(Mcl-1) . 3 Cell viability in different time after transfection was assayed By MTT and then drew the growth curve of each experimental group. The apoptosis of PANC-1 cells each experimental group was measured by flow cytometry in 48h after transfection.Results 1 After experimental exploration, the best transfection efficiency was 87.5%. 2 The Mcl-1 mRNA and protein of siRNA (Mcl-1) group were significantly downragulated (p <0.05). This effect reached a peak at 48h after transfection then declined. 3 Compared with the control group, liposome group and NC-siRNA group, PANC-1 cells proliferation rate of siRNA (Mcl-1) group obviously slowed down. 4 The apoptosis rates of experimental groups 48h after transfection were measurd by flow cytometry . The apoptosis rates of experimental groups were 14.6±0.36%( siRNA (Mcl-1) group), 4.1±0.35(control group), 4.2±0.41(liposome group), 4.4±0.26(NC-siRNA group),respectively. The apoptosis rate of siRNA (Mcl-1) group was significantly higher than other groups(p <0.05).Conclusions Liposome-mediated siRNA (Mcl-1) can efficiently inhibit the expression of Mcl-1 in PANC-1. The proliferation of PANC-1 cell decreased after downregulation of Mcl-1 and the apoptosis of PANC-1 cell increased after downregulation of Mcl-1. The RNAi targeting Mcl-1 can be seen a potential measure in pancreatic cancer.