茜草蒽醌诱导肝癌SMMC-7721细胞凋亡及其分子机制的研究

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。

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

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. G₁period of cell cycle had apoptosis peak of abnormal diploid by PI simple stain, and cell cycle stopped at G₁ period. The apoptosis fuction of anthraquinone introdution hepatoearcinoma 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.