目的：研究靶向下调小谷氨酰胺三角四肽重复区蛋白α抗体(small glutamine-rich tetratricopeptide repeat-containing protein alpha, SGTA)的表达对肝癌HuH7细胞周期及细胞增殖的影响。方法：选择HuH7细胞系作为研究对象，进行血清饥饿释放同步化处理后采用Western Blot方法检测SGTA蛋白表达水平变化；通过SGTA特异性siRNA下调HuH7细胞中SGTA的表达，采用细胞计数盒8(cell counting kit 8, CCK8)、流式细胞术等方法检测细胞增殖能力及细胞周期。结果：血清饥饿使HuH7细胞生长周期停滞，血清释放后刺激HuH7细胞增殖，Western Blot显示SGTA﹑增殖细胞核抗原(proliferating cell nuclear antigen, PCNA)蛋白随着细胞周期的进展表达增加，而细胞周期负性调控因子p27kip1蛋白表达却下降。 SGTA缺失可引起cyclin A、cyclin B的表达下降、细胞增殖的下降以及细胞周期的阻滞。结论：SGTA特异性siRNA能明显抑制肝癌HuH7细胞的增殖，细胞周期阻滞，提示其可为肝癌的靶基因治疗提供新的分子靶点。%Objective: To investigate the change of cell cycle and proliferation of hepatic cell carcinoma through targeted down-regulation the expression of small glutamine-rich tetratricopeptide repeat-containing protein alpha(SGTA) by the specific SGTA-siRNA. Methods: HuH7 cells were selected as the research object, Western Blot was used to detect SGTA protein expression level changes after serum starvation and refeeding, and analyze the role of SGTA in cell proliferation. Then, after knockdown of SGTA in HCC cells using small interfering RNAs, cell counting kit-8(CCK8) assay and flow cytometry were adopted respectively to determine cell proliferation and cell cycle progression in the cells. Results: The cell cycle of HuH7 cells was blocked by serum starvation, but the HuH7 cells were reentering the cell cycle after serum addition. Western Blot showed that the expression of SGTA and proliferatry cell nuclear antigen(PCNA) was increased, whereas the expression of p27 kip1 was inversely diminished after serum stimulation in HuH7 cells during cell cycle progression. siRNA analysis showed that SGTA depletion could inhibited cell proliferation and resulted in cell cycle arrest. Conclusions: siRNA-targeted SGTA could inhibit the cell proliferation and cell cycle of HuH7 cells which suggests that SGTA might serve as a new molecular target for the treatment of HCC.
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