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  

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

作　　者：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] 

机构地区：[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760190059

出　　处：《中国普通外科杂志》2016年第10期1431-1437,共7页China Journal of General Surgery

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（0-100mg／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；AnnexinVFIFC／PI双染法检测细胞的凋亡率；Westernblot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（0mg／LCu-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。Objective： To investigate the photodynamic killing effect colon cancer SW620 cells in vitro and the mechanism. mediated by Cu-Cy under X-ray activation on humanMethods： 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.

关 键 词：结肠肿瘤 光动力疗法 自噬 细胞凋亡 

分 类 号：R735.3[医药卫生—肿瘤]