Influence of RSK2 gene on the cell proliferation and apoptosis of human hepatocellular carcinoma cell line SMMC-7721
Farman Ali 1,2, HE Da-wei 3, HE Ao-lin 3, LI Sha-sha 3, ZHOU Ming-hui 3, WANG Bing-fang 2
(1. School of Medicine, Jiangsu University, Zhenjiang Jiangsu 212013; 2. Department of Gastroenterology, Kunshan Hospital Affiliated to Jiangsu University, Kunshan Jiangsu 215300; 3. Clinical Research & Lab Center, Kunshan Hospital Affiliated to Jiangsu University, Kunshan Jiangsu 215300, China)
Abstract:Objective: To investigate the effect of P90 ribosomal S6 kinase 2(RSK2) gene on the proliferation and apoptosis of human hepatocellular carcinoma cell line SMMC-7721. Methods: SMMC-7721 cells were divided into 5 groups, including control group, pcDNA3.1(+) vector group, pcDNA3.1(+)/RSK2 group, siRNAnegative control group and siRNA-RSK2 group. Control group was transfected with nothing, the other four groups were transfected with pcDNA3.1(+) empty plasmid, pcDNA3.1(+) /RSK2, siRNA- negative control and siRNA-RSK2, respectively. At 48 h post transfection, the RT-PCR and western blotting were respectively used to detect the expression of mRNA and protein of RSK2. CCK8 assay was used to measure the proliferation ability. And flow cytometry was used to detect apoptosis and cell cycle. Results: The results of RT-PCR and Western blotting showed that the mRNA and protein of RSK2 of pcDNA3.1(+)/RSK2 group were significantly higher than those of the other four groups(P<0.01). In contrast, the mRNA and protein of RSK2 of siRNA-RSK2 group were remarkably lower than those of the other four groups(P<0.01). The results of CCK8 assay indicated that overexpression of RSK2 gene promoted the proliferation of SMMC-7721 cells (P<0.01). Flow cytometry results showed that RSK2 gene silencing blocked the cell cycle in G1 phase(P<0.01), and decreased cell proliferation ability(P<0.01). The apoptosis rate of siRNA-RSK2 group was significantly higher than that of the other four groups(P<0.01), but the apoptosis rate of pcDNA3.1(+)/RSK2 group was not evidently different from that of the control group(P>0.05). Conclusion: Over-expression of RSK2 gene promoted the proliferation of SMMC-7721 cells, without any effect on cell apoptosis. The RSK2 gene silencing could inhibit the cell proliferation and promote the apoptosis.
[Key words]RSK2; hepatocellular carcinoma; cell proliferation; apoptosis