ARK5在肝癌组织中的表达及其对肝癌SMMC-7721细胞生长的影响

Expression of ARK5 in hepatocellular carcinoma tissue and its effect on growth of SMMC-7721 cells

目的:检测ARK5在肝癌组织及肝癌SMMC-7721细胞中表达水平,探讨其对肝癌细胞生长的作用。方法:采用PCR和Western blotting法检测30例肝癌组织及癌旁组织、正常肝细胞LO2及肝癌SMMC-7721细胞中ARK5 mRNA和蛋白表达水平;合成ARK5的小干扰RNA(siRNA)和阴性对照siRNA,转染至SMMC-7721细胞分别为实验组和阴性对照组,同时设空白对照组,采用MTT法和流式细胞术检测各组肝癌细胞的增殖活性和细胞凋亡率。结果:PCR和Western blotting法检测,肝癌组织和肝癌SMMC-7721细胞中ARK5mRNA和蛋白表达水平明显高于癌旁组织和LO2细胞(P<0.05)。MTT法检测,实验组24、48和72h细胞生长抑制率分别为(19.39±5.42)%、(23.19±0.53)%和(20.74±1.23)%,均高于空白对照组和阴性对照组(P<0.01);流式细胞术检测,实验组细胞凋亡率为(15.017±0.945)%,与空白对照组[(8.770±0.656)%]和阴性对照组[(8.763±1.201)%]比较差异均有统计学意义(P<0.05)。结论:ARK5在肝癌组织和肝癌SMMC-7721细胞中表达水平明显升高,抑制其表达可抑制肝癌细胞生长,促进肝癌细胞凋亡,提示ARK5可能作为一种癌基因促进肝癌的形成。

Objective To detect the expression of ARK5in hepatocellular carcinoma（HCC）tissue and hepatoma SMMC-7721cells,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 30cases of HCC tissue,paracarcinoma tissue,SMMC-7721cells,and hepatic cells LO2.The SiRNA of ARK5and negative control（NC）siRNA were constructed and transfected into the SMMC-7721cells,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 ARK5mRNA and protein in HCC tissue and SMMC-7721cells were significantly higher than those in paracarcinoma tissue and LO2cells（P〈0.05）.The MTT assay results demonstrated that the inhibitory rates of growth of transfected cells in experimental group at 24,48and 72hwere（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 differences compared with blank control group（8.770% ±0.656）% and negative control group（8.763% ±1.201%）（P〈0.05）.Conclusion The ARK5expression level is significantly increased in HCC tissue and hepatoma SMMC-7721cells;the inhibition of ARK5expression could suppress the growth of hepatoma cells and induce apoptosis.So ARK5maybe act as a cancer-promoting gene and induce hepatocellular carcinogenesis.