藏红花素诱导宫颈癌HeLa细胞凋亡及其作用机制  

Effect and mechanism of HeLa cell apoptosis induced by crocin

作　　者：龚青[1] 石丁波[1] 蔡卫斌[1] 杨中汉[1] 谭维格[1] 廖彩韵[1] 彭超[1] 刘畅[1] 高国全[1] 杨霞[1] 

机构地区：[1]中山大学中山医学院生物化学教研室,广东广州510080

出　　处：《中华肿瘤防治杂志》2009年第19期1445-1447,1462,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30600724;30700120;30872980);广州市科技计划项目(2008Z1-E231);中山大学医科2007年第一批学生业余科研立项项目

摘　　要：目的:研究藏红花素对HeLa细胞的促凋亡作用和机制。方法:设置0、0.2、0.4及0.8mmol/L4个浓度梯度,MTT法分析不同浓度藏红花素对HeLa细胞增殖的影响;设置空白阴性对照及秋水仙素(10μg/L)阳性对照,向HeLa细胞培养物分别加入0.4和0.8mmol/L的藏红花素,培养48h后用PI/AnnexinⅤ双染法分析其凋亡情况;1.6mmol/L藏红花素作用于HeLa细胞72h后,蛋白质印迹法检测Caspase-3的切割。结果:MTT实验分析显示,在0、0.2、0.4及0.8mmol/L浓度下HeLa细胞存活率分别为1、0.964、0.796和0.586。藏红花素显著地抑制HeLa细胞生长,P<0.05;各浓度组间均差异有统计学意义,P<0.05。PI/AnnexinⅤ双染法、流式细胞仪检测显示,阴性对照组、10μg/L秋水仙素阳性对照组、0.4及0.8mmol/L藏红花素处理组的细胞凋亡率分别为3.64%、15.99%、4.86%和9.28%,藏红花素促进HeLa细胞凋亡。蛋白质印迹法检测显示,藏红花素促进HeLa细胞Caspase-3切割。结论:藏红花素可明显地抑制HeLa细胞生长,并通过增加HeLa细胞Caspase-3切割促进其凋亡。OBJECTIVE：To investigate the growth inhibition and the apoptosis of HeLa cells induced by crocin,and explore the underlying molecular mechanism. METHODS：MTT essay was used to examine the suppressive effect of crocin on the proliferation of HeLa cells,at concentrations of 0,0.2,0.4 and 0.8 mmol/L. Other HeLa cells were cultured with normal medium or that mixed with colchicine （10 μg/L） or crocin （0.4 and 0.8 mmol/L） for 48 hours before they were harvested for examination by AnnexinⅤ and propidium iodide （PI） staining by the Annexin V-FITC Apoptosis Detection Kit. Western blot assay were used to determine the activation of Caspase-3 after 1.6 mmol/L crocin interacted with HeLa cells for 72 hours. RESULTS：MTT test analysis showed that in 0,0.2,0.4 and 0.8 mmol/L concentrations,the survival rates of HeLa cells were 1,0.964,0.796 and 0.586 respectively,and statistically significant differences existed among the different concentrations of crocin. Crocin significantly inhibited the growth of HeLa cells （P〈0.05）. AnnexinⅤ and propidium iodide （PI） staining showed that the apoptosis rates of HeLa cells of the PBS group,10 μg/L colchicine positive control group,0.4 mmol/L and 0.8 mmol/L crocin treated group were 3.64%,15.99%,4.86%,9.28%,respectively,indicating crocin promoted the apoptosis in HeLa cells. Caspase-3 was activated in the apoptosis of HeLa cells. CONCLUSION：Crocin can significantly inhibit the growth of HeLa cells through pro-apoptotic action,probably via the activation of Caspase-3.

关 键 词：藏红花苦素 HELA细胞 细胞凋亡 半胱氨酸天冬氨酸蛋白酶 

分 类 号：R737.33[医药卫生—肿瘤]