Selection of small interfering RNA targeting Glypican-3 and its influence on liver cancer cells

Abstract

Objective To study the action mechanism of Glypican-3 ( GPC3 ) in the development of hepatocellular carcinoma (HCC).Methods siRNA targeting GPC3 was designed and transfected into Huh7 cells,and the inhibitory effects on the expression of GPC3 was tested by using real-time polymerase chain reaction (PCR).The siRNA with obvious inhibitory effect was selected to down-regulate the GPC3 expression,and then the expression of GPC3 protein was detected by usig Wesnter blotting.The cell cycle,proliferation and invasion of Huh7 cells were assayed by using flow cytometry,fluorescence microscopy and Transwell,respectively.Results We successfully screened out a siRNA specifically targeting GPC3,its inhibition efficiency was 74.87％ and the siRNA could significantly inhibit the transcription and translation of GPC3.The knock-down of GPC3 could inhibit the proliferation and invasion of Huh7 cells,but had no effects on the cell cycle.Conclusion GPC3 expression facilitates the proliferation and invasion of HCC. Key words: Carcinoma,hepatocellular; Glypican-3; Small interfering RNA; Huh7 cells