基础研究(Basic Research)

X射线激发Cu-Cy介导的光动力对人结肠癌细胞的杀伤作用研究

Published at: 2016年第25卷第10期

刘志鹏 1 , 熊力 1 , 欧阳国庆 1 , 陈翔 1 , 周盼 1 , 王昆鹏 1 , 林良武 2 , 黄河 3 , 苗雄鹰 1 , 陈伟 4 , 文宇 1
1 中南大学湘雅二医院 普通外科,湖南 长沙,410011
2 中南大学粉末冶金学院,湖南 长沙,410083
3 中南大学湘雅医学院 组织学与胚胎学系,湖南 长沙,410078
4 美国德克萨斯州大学物理系,德克萨斯州 阿灵顿,76019-0059
通讯作者 宇 文 Email: 1850129046@qq.com
DOI: 10.3978/j.issn.1005-6947.10.3978/j.issn.1005-6947.2016.10.011
基金:
湖南省发改委课题基金资助项目 201583
中南大学中央高校专项资金资助项目 2016zzts537

摘要

目的:探讨X射线激发Cu-Cy介导的光动力对人结肠癌SW620细胞的体外杀伤效应及其作用机制。方法:用不同浓度(0~100 mg/L)Cu-Cy联合或不联合低剂量X线照射处理SW620细胞后,采用CCK8法检测并光镜观察细胞的生长活性;JC-1染色检测线粒体膜电位;AnnexinV FIFC/PI双染法检测细胞的凋亡率;Western blot检测凋亡相关蛋白Bax、Bcl-2及自噬相关蛋白LC3B、P62的表达;透射电镜观察细胞的超微结构。结果:与空白对照SW620细胞(0 mg/L Cu-Cy,无X线照射),Cu-Cy联合X线照射处理的SW620细胞生长活性明显降低,并呈一定的浓度依赖性(部分P<0.05),且细胞凋亡率明显升高、线粒体膜电位下降细胞的比例明显增加、Bax与LC3B-II蛋白表达明显升高,Bcl-2与P62蛋白表达明显降低(均P<0.05)、细胞内自噬体增多;单纯X射线照射和单加Cu-Cy处理的SW620细胞以上指标均无明显改变。结论:X线激发Cu-Cy介导的光动力能有效抑制人结肠癌SW620细胞体外生长活性,机制可能与其诱导细胞凋亡和自噬有关。


Photodynamic killing effect mediated by Cu-Cy under X-ray activation on colon cancer cells and the mechanism

Abstract

Objective: To investigate the photodynamic killing effect mediated by Cu-Cy under X-ray activation on human colon cancer SW620 cells in vitro and the mechanism. Methods: SW620 cells were exposed to different concentrations of Cu-Cy (0–100 mg/L) in combination with or without low dose of X-ray radiation, and then, the cell viability was examined by CCK8 assay and light microscopic observation, mitochondrial membrane potential was detected by JC-1 dye, apoptosis was measured by Annexin V/PI double-staining, the expressions of apoptosis-associated protein Bax and Bcl-2, and autophagy-associated protein LC3B and P62 were determined by Western blot analysis, and cell ultrastructure was observed by transmission electron microscopy. Results: In SW620 cells compared with blank control SW620 cells (without Cu-Cy treatment and X-ray radiation), the cell viability was significantly reduced with certain concentration-dependent manner (part P<0.05), and meanwhile, the ratio of cells with decreased mitochondrial membrane potential and the apoptotic rate were significantly increased, the expression levels of Bax and LC3B-II were significantly increased while the expression levels of Bcl-2 and P62 were significantly decreased (all P<0.05), and the intracellular autophagosomes were remarkably increased; all above indexes showed no significant change in those cells that underwent X-ray radiation alone or Cu-Cy treatment alone. Conclusion: The photodynamic action mediated by Cu-Cy under X-ray activation can effectively inhibit the growth and viability of human colon cancer SW620 cells in vitro, and the mechanism may possibly be associated with its inducing apoptosis and autophagy.


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引用

引用本文: 志鹏 刘, 力 熊, 国庆 欧阳, 翔 陈, 盼 周, 昆鹏 王, 良武 林, 河 黄, 雄鹰 苗, 伟 陈, 宇 文. X射线激发Cu-Cy介导的光动力对人结肠癌细胞的杀伤作用研究[J]. 中国普通外科杂志, 2016, 25(10): 1431-1437.
Cite this article as: LIU Zhipeng, XIONG Li, OUYANG Guoqing, CHEN Xiang, ZHOU Pan, WANG Kunpeng, LIN Liangwu, HUANG He, MIAO Xiongying, CHEN Wei, WEN Yu . Photodynamic killing effect mediated by Cu-Cy under X-ray activation on colon cancer cells and the mechanism[J]. Chin J Gen Surg, 2016, 25(10): 1431-1437.