醋酸棉酚诱导人粘液表皮样癌MEC-1细胞DNA双链断裂  

Gossypol acetic acid induces DNA double-strand breaks in human mucoepider-moid carcinoma cell MEC-1

作　　者：郭忠[1] 赵晋[1] 薛同敏[1] 马建秀[1] 汪晨净[1] 黄双盛[1] 

机构地区：[1]西北民族大学医学院,兰州730030

出　　处：《生理学报》2011年第2期164-170,共7页Acta Physiologica Sinica

基　　金：supported by the Projects for Young Scholars of Northwest University for Minorities;China(No.X2009-002/XB-MU-2010-BD-5)

摘　　要：本文研究醋酸棉酚(gossypol acetic acid,GAA)对人粘液表皮样癌细胞MEC-1体外增殖的影响,并初步探讨其抑制肿瘤细胞增殖的机制。体外培养人粘液表皮样癌细胞系MEC-1细胞,用MTT法检测GAA对MEC-1细胞增殖的影响;用中性彗星实验检测GAA对MEC-1细胞的DNA双链断裂;用免疫荧光染色法检测GAA诱导的磷酸化组蛋白γH2AX焦点形成。结果显示,5～40μmol/L的GAA以时间和浓度依赖方式抑制MEC-1细胞的生长;2.5～40μmol/L的GAA作用24h,或20μmol/L的GAA作用3～48h,CASP软件分析显示MEC-1彗星细胞的头部DNA百分含量减少,尾长、彗星长度、尾部DNA百分含量、尾矩和Olive尾矩增加;2.5～20μmol/L的GAA作用24h,或20μmol/L的GAA作用3～48h,γH2AX阳性细胞率随着浓度和时间的增加而增加。上述结果表明GAA抑制MEC-1细胞增殖,诱导DNA双链断裂是其抑制肿瘤细胞增殖的机制之一。The present study was conducted to investigate the effects of gossypol acetic acid（GAA） on the proliferation of human mucoepidermoid carcinoma cell line MEC-1 in vitro and its possible molecular mechanisms of DNA double-strand breaks（DSB）.MTT assay was performed to test the inhibition of proliferation of MEC-1 cells by GAA.DSB and γH2AX foci formation induced by GAA were detected by neutral comet assay and immunostaining.GAA（5-40 μmol/L） inhibited the growth of MEC-1 cells in a dose-and time-dependent manner.One of the indexes of comet assay,percentage of head DNA was decreased,however other indexes,in-cluding tail length,percentage of tail DNA,tail moment（TM） and Olive tail moment（OTM） were increased when treated with 2.5-40 μmol/L GAA for 24 h or 20 μmol/L GAA for 3-48 h,compared with those in control.The percentage of γH2AX-positive cells was also increased when MEC-1 was treated with 2.5-20 μmol/L GAA for 24 h or 20 μmol/L GAA for 3-48 h,compared with that in control.All these results show that GAA inhibits the proliferation of MEC-1,and DSB maybe one of the mechanisms of inhibitory effect of GAA on the growth of tumor cells.

关 键 词：醋酸棉酚 MEC-1 DNA双链断裂 γH2AX 

分 类 号：R730.2[医药卫生—肿瘤]