苦参碱诱导宫颈癌SiHa细胞凋亡的相关机制研究

Effects of Matrine on Apoptosis of Human Cervical Cancer Cell SiHa and Its Mechanism

目的:观察苦参碱体外诱导宫颈癌SiHa细胞凋亡效应并对其可能的作用机制进行初步探讨。方法:将不同浓度(0.00、0.25、0.50、1.00、2.00 mg/ml)的苦参碱分别作用于体外培养的宫颈癌细胞,在不同时间点(24、48、72 h),采用MTT法检测细胞的增殖情况;采用Annexin-Ⅴ和PI双染法在流式细胞仪上检测细胞的凋亡率;采用Western blot检测细胞中凋亡相关蛋白Caspase-3的表达。结果:各组苦参碱对SiHa细胞的增殖均有抑制作用,MTT和流式细胞结果显示,不同浓度的苦参碱对细胞生长抑制率与对照组相比存在明显差异(P<0.05),呈剂量效应正相关及时间效应正相关。加入Caspase-3抑制剂Z-DEVD-FMK后,各组细胞凋亡率与只加苦参碱组相比凋亡率下降。Western blot检测结果显示Caspase-3蛋白水平随苦参碱浓度的提高而增加(P<0.05)。结论:苦参碱可诱导宫颈癌SiHa细胞的增殖,促进其凋亡,其作用机制可能与提高Caspase-3活性有关。

Objective： To observe the role of matrine（Mat） induced apoptosis of human cervical cancer SiHa cells and its mechanism. Methods： SiHa cells were exposed to the concentrations of matrine 0.00, 0.25, 0.50, 1.00 and 2.00 mg/ml for 24, 48, 72 h. Cell proliferation was detected by MTT assay. Apoptosis was determined by double stained assay with Annexin- Ⅴand PI. Caspase-3 protein level was determined with western blot assay. Results： After SiHa cells were cocultured with matrine in vitro, the cell proliferation was inhibited in time and dose dependent manner. When the SiHa cells were cultured for 24, 48, 72 h by matrine 0.50, 1.00, 2.00 mg/ml, the total apoptosis rate was increased, which had ststistical significance compared with control group（P〈0.05）. The addition of the caspase-3inhibitor Z-DEVD-FMK, the apoptotic rate and increase of matrine group decreased apoptosis rate. And the caspase-3 protein level increased with the increase of the concentration of matrine（P〈0.05）. Conclusion： Matrine can inhibit the proliferation and induce the apoptosis of SiHa cells in vitro, one of its possible mechanisms is that matrine increasing caspase-3 activity.