Effects of RNAi-mediated gene silencing of LRIG3 expression on the cell cycle and apoptosls of glioma cells

Abstract

Objective To explore the effects of inhibiting the expression of LRIG3 gene on prolif-eration and apoptosis of human glioma cell line GL15 and explore its possible mechanism by using small interfering RNA (siRNA) targeting LRIG3 gene.Methods Two pairs of pGenesil2-LRIG3-shRNA (siR-NA) were transfected into GL15 glioma cells by Metafectine as experimental groups 1 and 2,and the cells that had a stably suppressive LRIG3 expression were selected by G418.The control ceils were transfected with negative shRNA.The changes in LRIG3 mRNA and protein levels were measured by RT-PCR and Western blot.The apoptosis rate and cell cycle were analyzed by flow cytometry.Results Compared with LRIG3 mRNA expression in the negative shRNA-treated GL15 cells, the transcription after treatment with LRIG3-specific shRNA was silenced by 52.4% and 63.8%,and the expression of LRIG3 protein was re-duced by about 50.9% and 67.4% in experimental groups 1 and 2 respectively.Cell cycle analysis showed that silencing LRIG3 increased the percentage of G2/M phase cells and improved the proliferation index significantly (P<0.01).Conclusion The siRNA targeting LRIG3 gene can dramatically inhibit RNA transcription and protein expression, then promote the proliferation of GL15 ceils and suppress the apoptosis of GL15 cells. Key words: Glioma; RNA interfering; Cell cycle; Apoptosis