葫芦素E对人乳腺癌细胞抑制作用及其机制的探讨  

Inhibitory effect of cucurbitacin E on the proliferation of breast cancer cells and its mechanism

作　　者：蓝天[1] 徐谦[2] 刘卫国[2] 金洪传[3] 毛伟敏[2] 王晓稼[2] 

机构地区：[1]杭州市中医院乳腺病科,浙江杭州310007 [2]浙江省肿瘤医院肿瘤内科浙江省中西医结合肿瘤重点实验室,浙江杭州310022 [3]浙江大学医学院附属邵逸夫医院生物医学研究中心,浙江杭州310003

出　　处：《中华肿瘤防治杂志》2013年第14期1065-1069,共5页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:研究葫芦素E(CuE)体外对人乳腺癌Bcap-37和MDA-MB-231细胞增殖的影响及其相关分子机制。方法:采用MTT法观察不同浓度CuE(0、0.1、1.0、10.0和100.0μmol/L)作用不同时间(24、48和72h)后对乳腺癌Bcap-37和MDA-MB-231细胞的影响;流式细胞仪检测CuE对乳腺癌细胞周期分布和细胞凋亡的影响;用蛋白质印迹法测定CuE处理后乳腺癌细胞内Pro-Caspase-3、cleaved Caspase-3及cleaved PARP蛋白水平的变化。结果:MTT结果显示CuE可显著抑制乳腺癌细胞的增殖,并呈时间和浓度依赖性,P<0.05。在CuE(0.1、1.0、10.0和100.0μmol/L)作用24h后,Bcap-37和MDA-MB-231细胞增殖抑制范围分别为17%～44%和12%～56%;而在作用48和72h后Bcap-37和MDA-MB-231细胞抑制率分别为24%～87%、36%～89%和33%～89%、61%～94%。流式细胞仪检测显示,CuE诱导乳腺癌细胞G2/M期阻滞和细胞凋亡。荧光显微镜显示,CuE处理组乳腺癌细胞出现典型的细胞核固缩现象。蛋白质印迹法结果显示,经CuE处理后乳腺癌细胞内cleaved Caspase-3及cleaved PARP蛋白水平上调,而Pro-Caspase-3水平则下降。结论:CuE可抑制乳腺癌细胞Bcap-37和MDA-MB-231细胞的增殖,并呈CuE浓度和时间依赖性,其抑制作用可能与诱导细胞凋亡和G2/M周期阻滞有关。OBJECTIVE： The present study was to observe the effect of Cucurbitacin E（CuE） on proliferation of breast cancer cells in vitrc and to explore corresponding molecular mechanism. METHODS： The inhibitory effect of varying concentration of CuE （0,0.1,1.0,10.0 and 100.0 mol/L） on proliferation of Bcap-37 and MDA-MB-231 cells for 24,48,72 hours was assessed by MTT assay; the changes of cell cycle distribution and cell apoptosis were determined by flow cytometry（FCM）; the expression of Pro-Caspase-3, cleaved Caspase-3 and cleaved PARP wa.s detected in breast cancer cells treated by Western Blot assay. RESULTS.. CuE inhibited breast cancer cellular proliferation in a dose and time- dependent manner （P〈0.05）. Reduction of Bcap-37, MDA-MB-231 cell viability with different concentration of CuE treatment （0.1,1.0,10.0 and 100.0 mol/L） after 24 hours ranged from 17 -- 44 , 12 M -- 56 % respectively, Whereas after 48,72 hours Bcap-37 and MDA-MB-231ranged from 24%--87 ,36--89M and 33%--89./0,61--94% respec- tively. As shown by FCM.CuE treatment （0,0. 1,1 mol/L for 24 h） of Bcap-37 and MDA-MB-231 resulted in G2/M phase arrest. FCM analysi;s also showed that CuE induced cell apoptosis and G2/M cell cycle arrest. The marked morphological changes of cell apoptosis were clearly observed in breast cancer cells treated with CuE. Cleaved caspase3 and cleaved PARP were upregalated and Pro-Caspase-3 protein was downregulated after CuE treatment. CONCLUSIONS.. CuE treatment resulted in a significant dose-dependent and time-dependent inhibition in growth of Bcap-37 and MDA-MB-231. CuE inhibits the proliferation of breast cancer cells through induction of G2/M phase arrest and cell apoptosis.

关 键 词：葫芦素E 乳腺肿瘤 凋亡 印迹法 蛋白质 

分 类 号：R737.9[医药卫生—肿瘤]