Hsa_circ_0000711在人肝癌细胞系中的功能研究

目的 研究环状RNA hsa_circ_0000711在人肝癌细胞系中的表达以及对肝癌细胞生物学功能的影响.方法 利用RT-qPCR检测环状RNA hsa_circ_0000711在肝癌细胞系的表达;通过小干扰RNA特异性敲低hsa_circ_0000711的表达;通过CCK-8实验检测肝癌细胞系的增殖情况;利用流式细胞技术检测细胞周期及细胞凋亡.Western blot检测下调hsa _circ _0000711后Cyclin D1、CDK4、 cleavaed Caspase 3及Bcl-2蛋白的表达水平.结果 Hsa_circ_0000711在肝癌细胞HepG2和SMMC- 7721中明显高表达(P＜0. 05) .与对照组相比,hsa_circ_0000711干扰组的肝癌细胞增殖能力显著降低(P＜0. 05),处于G1期细胞增多(P＜0. 05),且凋亡细胞显著增加(P＜0. 05) .干扰hsa_circ_0000711的表达后,肝癌细胞中Cyclin D1、CDK4和Bcl-2的蛋白表达量显著降低(P＜0. 05),cleavaed Caspase 3蛋白表达量显著升高(P＜0. 05) .结论 Hsa_circ_0000711可能通过调控细胞周期及凋亡影响肝癌细胞的增殖.

Function of circular RNA hsa_circ_0000711 in hepatocellular carcinoma cell lines

Objective To explore the expression profile of circular RNA (circRNA), hsa _circ _ 0000711, in hepatocellular carcinoma (HCC) cell lines and determine the effect of hsa_circ_0000711 on the biological behaviors of HCC cells. Methods The expression level of Hsa_circ_0000711 were detected by real-time polymerase chain reaction assay (RT-qPCR) in HCC cell lines HepG2 and SMMC-7721 and human live cell line L02. Small RNA interference was used to knock down the expression of hsa_circ_0000711. Then CCK-8 assay was employed to detect the proliferation, and flow cytometry for cell cycle and apoptosis. The protein levels of Cyclin D1, CDK4, cleaved Caspase 3 and Bcl-2 were measured by Western blotting. Results Hsa_circ_0000711 was more strongly expressed in the HepG2 and SMMC-7721 cells than the L02 cells(P＜0.05). Down-regulation of hsa_circ _0000711 significantly suppressed the growth rate of HepG2 and SMMC-7721 cells (P＜0.05), arrested the cells at cycle G1(P＜0.05), and promoted cell apoptosis (P＜0.05). After hsa_circ_0000711 interference, the protein expression levels of Cyclin D1, CDK4 and Bcl-2 were decreased notably (P＜0.05), but that of cleaved Caspase 3 was increased (P＜0.05). Conclusion Hsa_circ_0000711 may affect HCC cell proliferation by regulating cell cycle and cell apoptosis.