Lin X, Pan X M, Peng Z K, Wang K, Tang N
The Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), Chongqing 400016, China.
Chongqing Yucai Middle School, Chongqing 400050, China.
Zhonghua Gan Zang Bing Za Zhi. 2022 Feb 20;30(2):213-219. doi: 10.3760/cma.j.cn501113-20210204-00067.
To investigate the effects of glucose-6-phosphatase catalytic subunit (G6PC) recombinant adenovirus on proliferation and cell cycle regulation of liver cancer cells. 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. 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 ( < 0.05). Clone formation assay showed that the number of clones was significantly lower in AdG6PC than the other two groups ( < 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 ( < 0.05). Scratch repair rate was significantly lower in AdG6PC than the other two groups ( < 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. G6PC inhibits the proliferation, cell-cycle related expression, and migration of liver cancer cells by inhibiting the G(1)/S phase transition.
探讨葡萄糖-6-磷酸酶催化亚基（G6PC）重组腺病毒对肝癌细胞增殖及细胞周期调控的影响。构建重组腺病毒AdG6PC。将Huh7细胞和SK-Hep1细胞分为Mock组、AdGFP组和AdG6PC组。采用细胞增殖和克隆形成实验观察肝癌细胞的增殖情况。采用Transwell实验和划痕实验观察肝癌细胞的侵袭和迁移能力。采用细胞周期流式细胞术分析G6PC过表达对肝癌细胞增殖周期的影响。采用蛋白质免疫印迹法检测G6PC过表达对肝癌细胞周期相关蛋白表达的影响。成功构建了重组腺病毒AdG6PC。Huh7和SK-Hep1细胞增殖实验结果显示，AdG6PC组增殖细胞数明显低于其他两组（P<0.05）。克隆形成实验结果显示，AdG6PC组克隆数明显低于其他两组（P<0.05），提示G6PC过表达可显著抑制肝癌细胞的增殖。Transwell实验结果显示，AdG6PC组细胞迁移数明显低于其他两组（P<0.05）。划痕修复率AdG6PC组明显低于其他两组（P<0.05），提示G6PC过表达可显著抑制肝癌细胞的侵袭和迁移。细胞周期流式细胞术结果显示，G6PC过表达显著抑制Huh7细胞G(1)/S期转换。蛋白质免疫印迹结果显示，G6PC过表达下调了细胞周期相关蛋白的表达。G6PC通过抑制G(1)/S期转换抑制肝癌细胞的增殖、细胞周期相关表达及迁移。
