关键词: 肝肿瘤, RNA干扰, ARK5基因, SMMC-7721细胞

Abstract:

Abstract:Objective
To detect the expression of ARK5 in hepatocellular carcinoma(HCC) tissue and  hepatoma SMMC-7721 cells,and to investigate its effect on the growth of hepatoma cells.Methods The expression levels of ARK5 mRNA and protein were determined by RT-PCR and Western blotting in 30 cases of HCC tissue,paracarcinoma tissue,SMMC-7721 cells, and hepatic cells LO2.The SiRNA of ARK5 and negative control (NC) siRNA were constructed and transfected into the  SMMC-7721 cells,and used as experimental group and negative control group;at the same time blank control group was set up.The proliferation activity and apoptotic rate of transfected cells were detected by MTT assay and flow cytometry (FCM). Results The PCR and Western blotting results showed that the expression levels of ARK5 mRNA and protein in HCC tissue and SMMC-7721 cells were significantly higher than those in paracarcinoma tissue and LO2 cells(P＜0.05).The MTT assay results demonstrated that the inhibitory rates of growth of  transfected cells in experimental group at 24,48 and 72 h were (19.39±5.42)%,(23.19±0.53)%,and (20.74±1.23)%;there were significant differences compared with blank control group and negative control group (P＜0.01). The FCM results indicated that the apoptotic rate of the transfected cells in experimental group was (15.017±0.945)%,there were significant  ifferences compared with blank control group (8.770%±0.656)% and negative control group (8.763%±1.201%)(P＜0.05).Conclusion The ARK5 expression level is significantly increased in HCC tissue and hepatoma SMMC-7721 cells;the inhibition of ARK5 expression could suppress the growth of hepatoma cells and induce apoptosis.So ARK5 maybe act as a cancer-promoting gene and induce hepatocellular carcinogenesis.

Key words: liveer neoplasms, RNA interference, ARK5 gene, SMMC-7721 cells