The effect of Snail gene silencing on cell invasion and proliferation in human pancreatic cancer cell line PANC1 / 中华胰腺病杂志

ABSTRACT

Objective To observe the effect of silencing Snail gene on the invasion and proliferation ability of human pancreatic cancer cell line PANC1.Methods Lentiviral vectors that can express small hairpin RNA(shRNA) targeting human Snail gene(shRNA-Snail) or shRNA sequence that did not match any known mRNA(shRNA-NC) were constructed,and transfected into PANC1 cells.Untransfected cells served as control.mRNA and protein expression of Snail,α-smooth muscle actin (α-SMA) and E-cadherin was determined by real time quantitative PCR and Western blotting,respectively.In vitro invasion ability was tested by Transwell model.Proliferation ability was measured by CCK-8 assay.Results Compared with those in shRNA-NC group,Snail mRNA (0.27 ± 0.02 vs 0.92 ± 0.03) and protein level (0.26 ± 0.02 vs 0.80 ± 0.02),and α-SMA mRNA (0.33 ±0.04 vs 0.97 ±0.07) and protein level (0.31 ±0.04 vs 0.74 ±0.06) in shRNA-Snail group were obviously decreased,but E-cadherin mRNA (1.57 ± 0.45 vs 0.95 ± 0.08) and protein level (0.86 ± 0.03 vs 0.20 ± 0.03) were greatly increased.The number of cells permeating the septum of transwell [(6.80 ± 0.73)/400 magnification vs (26.80 ± 2.52)/400 magnification,P ＜0.01] was significantly decreased,and cell proliferation was inhibited (0.74 ± 0.05 vs 1.47 ± 0.04,P ＜ 0.01).All the differences above were statistically significant (all P ＜ 0.01).No significant differences were observed between shRNA-NC and normal control group.Conclusions Silencing Snail gene may restrain the invasion and proliferation ability of PANC1 cells to a certain degree.