Objective: To investigate the effects of glucose-6-phosphatase catalytic subunit (G6PC) recombinant adenovirus on proliferation and cell cycle regulation of liver cancer cells. Methods: Recombinant adenovirus AdG6PC was constructed. Huh7 cells and SK-Hep1 cells were set as Mock, AdGFP and AdG6PC group. Cell proliferation and clone formation assay were used to observe the proliferation of liver cancer cells. Transwell and scratch assay were used to observe the invasion and migration of liver cancer cells. Cell cycle flow cytometry assay was used to analyze the effect of G6PC overexpression on the proliferation cycle of liver cancer cells. Western blot was used to detect the effect of G6PC overexpression on the cell-cycle protein expression in liver cancer cells. Results: The recombinant adenovirus AdG6PC was successfully constructed. Huh7 and SK-Hep1 cells proliferation assay showed that the number of proliferating cells in the AdG6PC group was significantly lower than the other two groups (P < 0.05). Clone formation assay showed that the number of clones was significantly lower in AdG6PC than the other two groups (P < 0.05), suggesting that G6PC overexpression could significantly inhibit the proliferation of liver cancer cells. Transwell assay showed that the number of cell migration was significantly lower in AdG6PC than the other two groups (P < 0.05). Scratch repair rate was significantly lower in AdG6PC than the other two groups (P < 0.05), suggesting that G6PC overexpression can significantly inhibit the invasion and migration of liver cancer cells. Cell cycle flow cytometry showed that G6PC overexpression had significantly inhibited the Huh7 cells G(1)/S phase transition. Western blot result showed that G6PC overexpression had down-regulated the proliferation in cell-cycle related proteins expression. Conclusion: G6PC inhibits the proliferation, cell-cycle related expression, and migration of liver cancer cells by inhibiting the G(1)/S phase transition.
目的： 葡萄糖-6-磷酸酶催化亚基（G6PC）重组腺病毒，探究G6PC对肝癌细胞增殖及细胞周期调控的影响。 方法： 构建重组腺病毒AdG6PC,将Huh7细胞及SK-Hep1细胞设为Mock组、AdGFP组、AdG6PC组。采用细胞增殖实验及克隆形成实验观察其对肝癌细胞增殖的影响，transwell实验及划痕实验观察其对肝癌细胞侵袭和迁移的影响，细胞周期流式分析过表达G6PC对肝癌细胞增殖周期的影响，蛋白质印迹法检测过表达G6PC对肝癌细胞周期蛋白表达的影响。 结果： 成功构建重组腺病毒AdG6PC。Huh7细胞及SK-Hep1细胞增殖实验示AdG6PC组增殖细胞数明显低于其他2组（P < 0.05），克隆形成实验显示AdG6PC组的克隆数明显少于其他2组（P < 0.05），说明肝癌细胞中过表达G6PC可明显抑制肝癌细胞的增殖能力；transwell实验结果表明细胞AdG6PC组细胞迁移数明显少于其他2组（P < 0.05），划痕实验结果显示AdG6PC组划痕修复率明显低于其他2组（P < 0.05），说明过表达G6PC可明显抑制肝癌细胞侵袭和迁移。细胞周期流式分析表明，过表达G6PC显著抑制huh7细胞的G(1)/S期的转变。蛋白质印迹法结果示过表达G6PC可下调细胞增殖周期相关蛋白的表达。 结论： G6PC通过抑制肝癌细胞周期G(1)/S期转换及相关周期蛋白的表达抑制肝癌细胞增殖，同时G6PC也可抑制肝癌细胞迁移。.
Keywords: Cell cycle; Cell proliferation; Glucose-6 phosphatase catalytic subunit; Hepatocellular carcinoma.