醋酸棉酚诱导三株宫颈癌细胞DNA双链断裂研究

Research of Gossypol Acetic Acid Inducing DNA Double-Strand Breaks in Three Cervical Cancer Cell Lines

DOI：10.3969/j.issn.1673-1689.2018.02.002

中文关键词: 醋酸棉酚 宫颈癌 DNA双链损伤 γH2AX

英文关键词: gossypol acetate,cervical cancer,DNA damage,γH2AX

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中文摘要:

本文作者研究醋酸棉酚（ gossypol acetic acid，GAA）对3株宫颈癌细胞Hela、Siha、C-33A的DNA双链断裂（double-strand break，DSB）机制。MTT法检测GAA对3株宫颈癌细胞的体外增殖的影响；单细胞凝胶电泳实验检测GAA对3株宫颈癌细胞造成的DNA双链断裂；免疫组织化学法检测GAA诱导3株宫颈癌细胞产生的磷酸化组蛋白γH2AX焦点。GAA（0~160 μmol/L）以时间和浓度依赖方式抑制3株宫颈癌细胞的生长。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，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；而免疫组织化学染色结果显示，γH2AX阳性细胞率呈浓度、时间依赖性增加。3株细胞对GAA的敏感性呈现一定差异性。GAA抗肿瘤的作用机制为诱导细胞DSB，其中GAA诱导C-33A的DSB作用最强，Siha次之，Hela最弱。

英文摘要:

To explore the anti-tumor effect of three cervical cancer cell lines Hela，Siha and C-33A induced by gossypol acetic acid（GAA） respectively and the mechanism. The inhibitory effect of GAA on the growth of three cervical cancer cell lines was detected by MTT assay.The DNA damage of three cervical cancer cell lines caused by GAA was detected by single cell gel electrophoresis. The expression of phosphorylated histone protein（γH2AX） in three cervical cancer cell lines produced by GAA was detected by immunohistochemistry. Results：GAA（0~160 μmol/L） inhibited the growth of three cervical cancer cell lines in a dose-time dependence. The inhibition rate increased with the increase of concentration and time. 2.5~40 μmol/L of the GAA for 24 h，or 20 μmol/L GAA role 3~48 h，The result of single cell gel electrophoresis display that cell comet head DNA percentage was reduced，but tail length，comet length，tail DNA percentage，tail moment and olive tail moment was increased；and immunohistochemistry results showed that，the γH2AX positive rate increased in a concentration-time dependence. The mechanism of antitumor by GAA is to inhibit the proliferation of three cervical cancer cell lines，induce DNA double strand breaks，and lead to cell death. And the sensitivity of C-33A was the highest，the Hela was the second，and Siha was the most insensitive.

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