槲皮素联合survivin反义核苷酸对肝癌细胞凋亡协同作用的研究

目的 探讨槲皮素联合survivin反义核苷酸(ASODN)对肝癌SSMC-7721细胞株增殖、凋亡和细胞周期的影响.方法 常规培养SSMC-7721细胞,用四甲基偶氮唑盐法(MTT法)评价survivin ASODN联合槲皮素对肝癌细胞增殖的影响;流式细胞仪(FCM)检测细胞凋亡率和细胞周期;荧光染色观察细胞形态学变化;并通过RT-PCR和免疫组化方法检测survivin基因表达变化.结果 survivin反义寡核苷酸转染SSMC-7721细胞后,可以显著抑制细胞增殖,其抑制作用具有剂量依赖性,且能诱导肝癌细胞凋亡;联合槲皮索和survivin反义寡核苷酸抑制作用更为显著(t=4.317,P<0.01);RT-PCR及免疫组化显示ASODN和槲皮素均使survivin mRNA和蛋白表达下降.结论 survivin基因反义寡核苷酸联合槲皮素能明显抑制肝癌SSMC-7721细胞增殖、诱导细胞凋亡和下调survivin基因表达;两者具有协同作用.

The effect of survivin antisense oligodeoxynucleotides with quercetin on HCC cell lines

Objective To study the effect of survivin anfisense oligonucleotides (ASODN)combined with quercetin on proliferation, apoptosis and cell cycle of a hepatocellular carcinoma cell line SSMC-7721 cells. Methods Human hepatocellular carcinoma cell line SSMC-7721 was cultured in vitro,and cells on logarithmic growth phase were used for this experiment. Cell proliferation was measured by MTT assay. The apoptotic rate and cell cycle were examined by flow cytometer (FCM). Morphological change of apoptotic cells were observed by fluorescent microscope. The expression of survivin gene was detected by the method of immunohistochemistry staining and RT-PCR on the mRNA and protein level. Results After sealing survivin gene with ASODN, the proliferation of SSMC-7721 cells was inhibited markedly. FCM analysis showed that there appeared an obvious apoptosis peak after transfection. The inhibitory effect of combined administration of survivin ASODN and quercetin on cell proliferation was much stronger than that of the single way. The result of immunohistochemical and RT-PCR assays showed that survivin ASODN and quercetin inhibited the expression of survivin gene. Conclusion Combined survivin ASODN with quercetin significantly inhibit cell proliferation, down-regulate survivin gene expression and induce the apoptosis of SSMC-7721 cells.