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醋酸棉酚诱导人粘液表皮样癌MEC-1细胞DNA双链断裂

郭忠, 赵晋^*, 薛同敏, 马建秀, 汪晨净, 黄双盛

西北民族大学医学院，兰州 730030

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，5~40 μmol/L的GAA以时间和浓度依赖方式抑制MEC-1细胞的生长；2.5~40 μmol/L的GAA作用24 h，或20 μmol/L的GAA作用3~48 h，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；2.5~20 μmol/L的GAA作用24 h，或20 μmol/L的GAA作用3~48 h，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。

关键词： 醋酸棉酚; MEC-1; DNA双链断裂; γH2AX

分类号：R737

[Gossypol acetic acid induces DNA double-strand breaks in human mucoepidermoid carcinoma cell MEC-1.] [Article in Chinese]

Guo Zhong, ZHAO Jin^*, XUE Tong-Min, MA Jian-Xiu, WANG Chen-Jing, HUANG Shuang-Sheng

Medical College of Northwest University for Nationalities, Lanzhou 730030, China

Abstract

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, including 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.

Key words: gossypol acetic acid; human mucoepidermoid carcinoma cell-1; double-strand breaks; γH2AX

收稿日期：2010-10-27　　录用日期：2011-01-07

通讯作者：赵晋　　E-mail: yxzj@xbmu.edu.cn

引用本文：

郭忠, 赵晋, 薛同敏, 马建秀, 汪晨净, 黄双盛. 醋酸棉酚诱导人粘液表皮样癌MEC-1细胞DNA双链断裂[J]. 生理学报 2011; 63 (2): 164-170.

Guo Zhong, ZHAO Jin, XUE Tong-Min, MA Jian-Xiu, WANG Chen-Jing, HUANG Shuang-Sheng. [Gossypol acetic acid induces DNA double-strand breaks in human mucoepidermoid carcinoma cell MEC-1.] [Article in Chinese]. Acta Physiol Sin 2011; 63 (2): 164-170 (in Chinese with English abstract).