木犀草素诱导结肠癌HT-29细胞凋亡及其作用机制  

Luteolin-induced apoptosis of colorectal carcinoma HT-29 cells and mechanism

作　　者：陈刚[1] 谢从华[1] 张俊红[1] 

机构地区：[1]武汉大学中南医院放化疗科,430071

出　　处：《中华实验外科杂志》2015年第11期2705-2707,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 探讨木犀草素在体外诱导结肠癌HT-29细胞凋亡的作用机制.方法 体外培养人结肠癌细胞HT-29,以不同浓度木犀草素处理后,应用噻唑蓝（MTT）法检测木犀草素对HT-29细胞增殖的抑制作用;膜联蛋白V-异硫氰酸荧光素（Annexin V-FITC）/碘化丙锭（PI）双标记流式细胞术检测细胞凋亡率;采用实时定量聚合酶链反应（Real-time PCR）检测B细胞淋巴瘤/白血病-2（bcl-2）、B细胞淋巴瘤/白血病-2相关X蛋白（bax）、半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3、Caspase-9 mRNA表达水平;Western blot检测木犀草素对HT-29细胞内bcl-2、bax、Caspase-3、Caspase-9等蛋白表达的影响.结果 MTT结果显示木犀草素能显著抑制HT-29细胞的生长、增殖,呈浓度和时间效应关系（72 h最高抑制率达78.72％）;流式细胞仪分析示木犀草素能诱导细胞发生凋亡,凋亡率可达（23.78±1.03）％;Real-time PCR检测结果示木犀草素上调了bax、Caspase-3、Caspase-9 mRNA表达,抑制了bcl-2 mRNA表达;木犀草素干预后,Western blot结果示bcl-2蛋白呈浓度剂量依赖性递减;而bax、Caspase-9、Caspase-3蛋白表达则显著增高.结论 木犀草素能通过线粒体通路抑制人结肠癌HT-29细胞的增殖并诱导其凋亡.Objective To explore the action mechanism of Luteolin-induced apoptosis of human colorectal carcinoma HT-29 cells.Methods HT-29 cells were cultured in vitro, and treated with Luteolin at different concentrations.The inhibitory effects of Luteolin on cell proliferation was examined by methyl thiazol tetrazolium （MTT） assay, and the inhibition rate of 80 μmol/L Luteolin for 72 h was 78.72%.The cell apoptosis rate was detected by Annexin V-fluoresceine isothiocyanate （FITC）/propidium iodide （PI） double staining, and the apoptosis rate of HT-29 cells treated with Luteolin for 24 h was （23.78 ± 1.03） %.The mRNA and protein levels of B cell lymphoma/leukemia-2 associated X protein （bax）, B cell lymphoma/leukemia-2 （bcl-2）, Caspase-3 and Caspase-9 were detected by real-time quantitative polymerase chain reaction （Real-time PCR） and Western blotting respectively.Results MTT assay showed Luteolin could inhibit the growth and proliferation of HT-29 cells in a time-and concentration-dependent manner.Flow cytometry showed that Luteolin could induce apoptosis of HT-29 cells.Luteolin could activate the expression of bax and Caspase-3,-9, and inhibit the expression of bcl-2.Conclusion Luteolin can significantly inhibit the proliferation and induce the apoptosis of HT-29 cells, which may be associated with mitochondrial pathway.

关 键 词：木犀草素 结肠癌 线粒体 

分 类 号：R735.35[医药卫生—肿瘤]