茜草蒽醌诱导肝癌SMMC-7721细胞凋亡及其分子机制的研究

Study of introduction apoptosis of hepatocarcinoma cell SMMC-7721 by anthraquinone extracted from rubiqmnone and molecular mechanism

目的:探讨茜草提取物蒽醌单体对人肝癌SMMC-7721细胞生长的抑制作用,诱导凋亡作用及对凋亡相关基因bcl-2表达的影响,为肝癌的基因治疗提供有效靶点。方法:采用MTT法检测生长的抑制作用;以形态学,DNA凝胶电泳,流式细胞仪单染、双染检测细胞凋亡;以RT-PCR来分析蒽醌对bcl-2 mRNA的影响。结果:蒽醌能抑制肝癌细胞的生长;蒽醌处理肝癌细胞后,倒置显微镜和光镜下可见典型的凋亡细胞;DNA琼脂糖凝胶电泳显示出典型的细胞凋亡梯形带,PI单染表明细胞周期的G1期前有异常二倍体细胞的凋亡峰,并将细胞周期阻滞在G1期,Annexin V-FITC/PI定量检测进一步证实了蒽醌诱导肝癌细胞凋亡的作用;RT-PCR检测表明蒽醌可使bcl-2 mRNA表达水平下降。结论:蒽醌能显著抑制肝癌细胞的生长,并诱导肝癌细胞凋亡,其分子机制可能与下调bcl-2基因的表达有关。

Objective： To research the effect of anthraquinone extracted from rubiqmnone on growth inhibition , introduction apoptosis and expression of relative gene of bcl-2 of hepatocarcinoma cell SMMC 7721, and provide the effective target of gene therapy. Method： The growth inhibition effect was detected by MTT. Morpholgy, DNA agarose gel electrophoresis and flow cytometry were used to observe the cell apoptosis. The effect of anthraquinone on bcl-2mRNA expression was analyzed by RT-PCR. Result： Anthraquinone could inhibit the growth of hepatoearcinoma cell SMMC 7721. The typical apoptosis cells were found by inverted microscope and common microscope. DNA agarose gel electrophoresis showed a typical apoptosis strip. G1 period of cell cycle had apoptosis peak of abnormal diploid by PI simple stain, and cell cycle stopped at G1 period. The apoptosis ruction of anthraquinone introdution hepatoeareinoma cell was further certified by Annexin V-FITC/PI. Anthraquinone could decrease the expression of bcl-2mRNA by RT-PCR. Conclusion ： Anthraquinone can significantly inhibit growth of hepatoearcinoma cell and induce apoptosis. The mocular mechanism may be related to down-regulating the expression of bcl-2 gene.