The effect of silencing alpha - fetoprotein expression in HepG2 cell on the cellular function

Abstract

Objective To construct lentiviral vectors with small interfering RNA (siRNA) targeted alpha-fetoprotein (AFP) and investigate the effect of silencing AFP expression on proliferation, invasion and metastasis of HepG2 cells. Methods Synthetic siRNA-AFP and its negative control were transfected into HepG2 cell by lentiviral vector, then getting a stable cell line with AFP expression silenced by puromycin. Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) and Western blotting was used for detecting the effects of interference at mRNA and protein levels. In addition, methyl thiazol tetrazolium (MTT) assay was used to measure the proliferation of HepG2 cell. The invasion ability changes were analyzed by Transwell and the migration of HepG2 cell was detected by wound healing assay. Results The results of FQ-PCR and Western blotting demonstrated that the AFP expression was significantly decreased about 90% and (62.53±6.17)%, respectively both at mRNA and protein levels after transfection (P<0.05). The proliferation abilities of HepG2 cell were apparently decreased (31.17±1.16)% by silencing AFP expression after culturing 6 days (P<0.05). The inhibition of silencing AFP could induce suppression of invasion and migration (P<0.05). Conclusion Silencing of AFP expression can effectively inhibit proliferation, invasion and metastasis process in HepG2 cell. Key words: Alpha-fetoprotein; RNA interference; HepG2 cell; Invasion; Metastasis