目的: 探讨沉默胶质瘤相关癌基因同源蛋白2 基因（glioma associated oncogene homolog 2，Gli2)对肝癌SMMC-7721细胞增殖、凋亡的影响及其可能的作用机制。方法:以重组shRNA-Gli2 慢病毒感染SMMC-7721 细胞,筛选沉默效果最佳干扰序列。以携最佳干扰序列重组慢病毒感染SMMC-7721 细胞为干扰组，shRNA-NC慢病毒感染SMMC-7721 细胞为对照组，不作处理SMMC-7721 细胞为空白组。采用CCK-8 法与集落形成实验检测各组细胞的增殖活性，流式细胞术检测各组细胞凋亡率；用qPCR 和Western blotting 法检测各组细胞Gli2、cyclinD1、Bax 和Bcl-2 mRNA及其蛋白质的表达。结果:重组慢病毒感染率约为90%，重组shRNA-Gli2-1 慢病毒沉默效果最佳；干扰组SMMC-7721 细胞Gli2 mRNA和蛋白质表达显著低于对照组与空白组（均P<0.05），而对照组与空白组之间Gli2 mRNA和蛋白质表达差异不显著（P>0.05）。干扰组SMMC-7721 细胞的增殖和细胞集落形成数明显少于对照组和空白组（均P<0.05）；干扰组SMMC-7721 细胞凋亡率显著高于对照组和空白组（均P<0.01）；干扰组SMMC-7721 细胞cyclinD1 及Bax mRNA和蛋白质表达显著低于对照组和空白组，而Bcl-2 mRNA和蛋白质表达则显著高于对照组和空白组（均P<0.05）。结论: 沉默Gli2 基因的表达能够抑制肝癌SMMC-7721 细胞增殖，促进肝癌细胞凋亡，其机制可能与cyclinD1、Bax的下调和Bcl-2 上调有关。

Objective: To explore effect of silencing glioma associated oncogene homolog 2 (Gli2) on proliferation and apoptosis of the liver cancer SMMC-7721 cell and its possible action mechanism. Methods: Infection of the SMMC-7721 cell with recombinant shRNA-Gli2 lentivirus was used to screen the best jamming sequence for silence effect. The SMMC-7721 cell of interference group was infected with the best jamming sequence lentivirus,the SMMC-7721 cell of control group was infected with shRNA-NC shRNA-Gli2 lentivirus and the SMMC-7721 cell of blank group didn’t with any treatment. CCK-8 and Western blotting assays were used to detect proliferation activity of the SMMC-7721 cell in various groups. Flow cytometry assay was used to test apoptosis of the SMMC-7721 cell in various groups. Expressions of Cli2, cyclinD1, Bax and Bcl-2 mRNA and their proteins were detected by qPCR and Western blotting. Results: Infection rate of the recombinant lentivirus was about 90%. Silence effect of the recombinant shRNA-Gli2-1 lentivirus was the best. Expressions of Gli2 mRNA and its protein in the SMMC-7721 cell of interference group were remarkably lower than those in the SMMC-7721 cells of control and blank groups (all P<0.05), and differences of Gli2 mRNA and its protein expressions between the SMMC-7721 cells of control group and blank group were not significant (P>0.05). Proliferation and cell colony forming number of the SSMC-7721 cell in interference group were obviously less than those of the SSMC-7721 cell in control and blank groups (all P<0.05). Apoptosis rate of the SMMC-7721 cell in interference group evidently higher than those of the SMMC-7721 cell in control and blank groups (P<0.01). Expressions of cyclinD1 and Bax mRNA and their proteins in the SMMC-7721 cell of interference group were observably lower than those in the SMMC-7721 cells of control and blank groups (all P<0.05), but expressions of Bcl-2 mRNA and protein in the SMMC-7721 cell of interference group were observably higher than those in the SMMC-7721 cells of control and blank groups (all P<0.05). Conclusion:Silencing expression of Gli2 gene could inhibit growth of the liver cancer SMMC-7721 cell, and promote apoptosis of the liver cancer cell, its mechanism might relate to down-regulation of cyclin D1 and Bax as well as upregulation of Bcl-2.

四川省卫生计生委科技科研项目资助（No.17PJ590）