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目的研究Bcl 2基因家族促凋亡蛋白Bak在细胞凋亡细胞周期调控中的作用 ,评价它们作为肿瘤治疗的潜在靶基因的可能性。方法采用一种可诱导性表达系统,(MT II调节系统),在外加锌离子(ZnSO4,100μmol/L)的条件下诱导Bak基因表达。以Hela细胞系作为靶细胞 ,获得表达Bak基因的稳定转染子。结果可诱导性Bak基因过表达的Hela细胞出现广泛死亡。TUNEL染色证实细胞核碎片化 ,提示细胞死亡是凋亡。流式细胞仪显示 ,在诱导后24h有19.29 %的细胞发生凋亡且细胞在G1 期明显积聚。结论Bak基因能显著诱导Hela细胞凋亡和细胞周期在G1 期延长。因此 ,Bak基因可能成为肿瘤基因治疗的靶基因
Objective To investigate the role of the pro-apoptotic protein Bak of the Bcl 2 gene family in the regulation of cell cycle in apoptosis and to evaluate their potential as target genes for cancer therapy. Methods An inducible expression system (MT II regulation system) was used to induce Bak gene expression under ZnSO4 (100μmol / L). Using Hela cell lines as target cells, stable transfectants expressing Bak gene were obtained. As a result, Hela cells that were overexpressed in the Bak gene were found to be widespread. TUNEL staining confirmed nuclear fragmentation, suggesting that cell death is apoptosis. Flow cytometry showed that 19.29% of the cells were apoptotic at 24 h after induction and the cells accumulated significantly in G1 phase. Conclusion Bak gene can significantly induce Hela cell apoptosis and cell cycle prolongation in G1 phase. Therefore, Bak gene may become the target gene of tumor gene therapy