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Effect of singal molecules on apoptosis incuced by Genistein in hepatocarcinoma cells
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摘要: 目的通过检测三羟异黄酮诱导肝癌细胞凋亡过程中信号分子的表达,以期探讨其诱导凋亡的分子机制。方法应用体外细胞培养技术,三羟异黄酮处理后行HE染色;采用免疫细胞化学和RT-PCR分别检测癌细胞凋亡过程中信号分子的表达情况。结果（1）三羟异黄酮作用后可见凋亡细胞,随着浓度的增加,凋亡率逐渐升高;（2）survivin蛋白的表达下调,caspase-3的蛋白表达上调;（3）survivin基因的表达逐渐降低,而caspase-3在基因水平的表达有逐渐增加趋势,但无统计学差异（P>0.05）。结论三羟异黄酮能诱导人肝癌细胞的凋亡;在此凋亡过程中,信号分子survivin在基因与蛋白水平呈现一致性的表达降低,caspase-3在蛋白水平明显上调,但其基因表达无明显变化。推测其作用靶点可能是在转录后水平。Abstract: Objective To investigate the molecular mechanism of apoptosis induced by genistein in hepatocellular cells through detecing expressions of caspase-3 and survivin.Methods Human hepatocellular cells were treatec with different concentration of genistein, the morphology of apoptosis cells were observed by nicroscopy, the expressions of caspase-3 and survivin were identified by immuno-cytochem cal staining method and RT-PCR.Results Hepatocarcinoma cells present some typical morphologic features of apoptosis, The apoptotic rate was increased with concentration.The expression of survivin protein and gene were lower in treated groups, The expression of caspase-3 protein vas up regulated, while the mRNA expression of caspase-3 was not changed.Conclusion Genistein can induced apoptosis of hepatocarcinoma cells, which accombied by downregulation of survivm gene and upregulation of caspase-3 ptotein, the molecular event of cell apoptosis may be dependent on the activaton of caspase-3 enzyme rather than the expression of caspase-3 gene.
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Key words:
- hepatocarcinoma cells /
- apoptosis /
- singal molecules
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[1]Ravindranath MH, Muthugounder S, Presser N, et al. Anticancer therapeutic potential of soyisoflavone, genistein[J].Adv Exp Med Biol, 2004, 546∶121-165. [2]Su SJ, Chow NH, Kung ML, et al.Effects of soy isoflavones on ap-optosis indutio G2/M arrest in human hepatoma cells involvementcaspase-3 activation, Bcl-2 and Bcl-XL downregulation andCdc2 kinase activity[J].J Nutr Cancer, 2003, 45 (1) ∶113-123. [3]Blanc-Brude OP, Mesri M, Wall NR, et al.Therapeutic targetingof the survivin pathway in cancer:initiation of mitochondrial apopto-sis and suppression of tumor-associated angiogenesis[J].ClinCancer Res, 2003, 9∶2683-2692. [4]Ikeguchi M, Ueda T, Sakatani T, et al.Expression of survivin mes-senger RNAcorrelates with poor prognosis in patients with hepatocel-lular carcinoma[J].Diagn Mol Pathol, 2002, 11 (1) ∶30-40. [5]Chodon D, Banu SM, Padmavathi R, et al.Inhibition of cell prolif-eration and induction of apoptosis by genistein in experimental hepa-tocellular carcinoma[J].Mol Cell Biochem, 2006, 297 (1-2) ∶73-80. -
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