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目的:探讨Tg737基因过表达对人肝癌细胞株细胞周期和凋亡的影响及可能的机制。方法:将肝癌HepG2和MHCC97-H细胞分别用脂质体法转染pcDNA3.1-Tg737质粒(Tg737过表达组)或pcDNA3.1空载体(空载体组),或单纯脂质体孵育(脂质体组),以各自未处理的细胞为空白对照。48h后分别用流式细胞仪检测细胞周期与凋亡,Hoechst33342染料法检测细胞核形态,Westernblot法检测cyclinA、Bax、Bcl-2表达。结果:与各自的空白对照组比较,Tg737过表达组HepG2和MHCC97-H细胞的S期细胞数量与细胞凋亡率均明显增加(均P<0.05),且细胞核均出现明显凋亡形态学改变,同时伴随cyclinA、Bax的表达上调和Bcl-2的下调(均P<0.05);空载体组、脂质体组HepG2和MHCC97-H细胞镜下未见明显的凋亡形态学变化,且上述指标差异均无统计学意义(均P>0.05)。结论:Tg737基因过表达可以抑制肝癌细胞周期进程、促进其凋亡,机制可能与Tg737参与调节cyclinA、Bax、Bcl-2有关的信号通路有关。
Objective: To investigate the effect of Tg737 gene overexpression on the cell cycle and apoptosis of human hepatoma cell line and its possible mechanism. Methods: HepG2 and MHCC97-H cells were transfected with pcDNA3.1-Tg737 plasmid (Tg737 overexpression group) or pcDNA3.1 empty vector (empty vector group) by lipofectamine or lipofectamine Plastid group), with untreated cells as a blank control. After 48h, the cell cycle and apoptosis were detected by flow cytometry. The nuclear morphology was detected by Hoechst33342 dye method. The expressions of cyclinA, Bax and Bcl-2 were detected by Western blot. Results: Compared with the blank control group, the number of S phase cells and the apoptosis rate of HepG2 and MHCC97-H cells in Tg737 over-expression group were significantly increased (all P <0.05), and obvious apoptotic morphological changes appeared in the nucleus , Accompanied by up-regulation of cyclin A and Bax and down-regulation of Bcl-2 (all P <0.05). No obvious apoptotic morphological changes were observed in HepG2 and MHCC97-H cells in empty vector group, There was no significant difference between the indexes (all P> 0.05). CONCLUSION: Tg737 overexpression can inhibit cell cycle progression and promote apoptosis of hepatocellular carcinoma cells. The mechanism may be related to the involvement of Tg737 in the regulation of cyclin A, Bax and Bcl-2 signaling pathways.