苦参碱诱导宫颈癌siha细胞凋亡的机制研究.doc

苦参碱诱导宫颈癌SiHa细胞凋亡的相关机制研究* 周美英1 魏林珍1 王海琳1# 张爱笠2 甄洁玉1 （1甘肃省人民医院妇产科 兰州730000；2甘肃中医药大学 兰州730000） 摘要：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。 关键词：苦参碱；宫颈癌；细胞增殖；细胞凋亡；Z-DEVD-FMK；Caspase-3 Effects of Matrine on Apoptosis of Human Cervical Cancer Cell Line SiHa and Its Mechanism ZHOU Mei-ying1, WEI Lin-zhen1, WANG Hai-lin1#, ZHANG Ai-li2, ZHEN Jie-yu1 (1Department of Obstetrics and Gynecology, Gansu Provincial Hospital, Lanzhou730000; 2Gansu University of Traditional Chinese Medicine, Lanzhou730000) Abstract: Objective: To invest 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, 0.25, 0.5, 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-Ⅴ and 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 culture for 24, 48, 72 h by matrine 0.5, 1.0, 2.0 mg/ml, the total apoptosis rate was increased, which had ststistical significance compared with control group (P all 0.05), After treatment with matrine (0.5, 1.0, 2.0 mg/ml) in the presence (+) or absence (-) of Z-DEVD-FMK (25 umol/L) for 48 h, SiHa cells were partially blocked by caspase-3 inhibitor Z-DEVD-FMK. The protein expression level of Caspase-3 increased after SiHa cells were treated by matrine 0, 0.5, 1.0, 2.0 mg/ml wit