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[目的]研究苦参碱(matrine,MA)对体外诱导人结肠癌HT29细胞的凋亡作用及其机制。[方法]不同浓度MA(0~32mg/ml)作用HT29细胞24h后,流式细胞仪检测细胞凋亡,线粒体跨膜电位检测试剂盒(JC-1)检测线粒体膜电位(Δφm)变化,分光光度法检测caspase-3,-9酶活性,Western blot法检测线粒体凋亡途径相关蛋白Bax、Bcl-2表达。[结果]MA显著诱导HT29细胞凋亡;16mg/ml MA作用HT29细胞24h后,caspase-9,-3酶活性明显升高;Western blot结果显示:MA处理组促凋亡蛋白Bax表达明显升高,抗凋亡蛋白Bcl-2表达明显降低。[结论]MA具有促进结肠癌细胞凋亡的作用,且具有剂量依赖性,其机制与线粒体凋亡途径有关。
[Objective] To investigate the apoptosis effect of matrine (MA) on human colon cancer HT29 cells in vitro and its mechanism. [Method] The apoptosis of HT29 cells was detected by flow cytometry after different concentrations of MA (0 ~ 32 mg / ml) for 24 h. The changes of mitochondrial membrane potential (Δφm) were detected by mitochondrial transmembrane potential detection kit (JC-1) The activity of caspase-3 and -9 were detected by spectrophotometry. The expressions of Bax and Bcl-2 in mitochondrial apoptosis pathway were detected by Western blot. [Results] MA significantly induced apoptosis of HT29 cells; the activity of caspase-9 and -3 in HT29 cells treated with 16 mg / ml MA for 24 h was significantly increased; Western blot results showed that the expression of Bax protein in MA-treated group was significantly increased , Anti-apoptotic protein Bcl-2 expression was significantly reduced. [Conclusion] MA can promote the apoptosis of colon cancer cells in a dose-dependent manner. The mechanism is related to mitochondrial apoptosis pathway.