原卟啉介导的光动力疗法对结肠癌HCT116细胞的杀伤和促凋亡作用及其机制  

Role of PpIX-based photodynamic therapy in promoting the damage and apoptosis of colorectal cancer cell and its mechanisms

作　　者：欧阳国庆[1] 刘志鹏[1] 熊力[1] 陈翔[1] 李清龙[1] 黄河[2] 林良武[3] 苗雄鹰[1] 马伦 陈伟[1,4] 文宇[1] OUYANG Guoqing LIU Zhipeng XIONG Li CHEN Xiang LI Qinglong HUANG He LIN Liangwu MIAO Xiongying MA Lun CHEN Wei WEN Yu(Department of General Surgery, Seeond Xiangya Hospital, Central South University, Changsha 41001 I Department of Histology and Embryology, Xiangya School of Medicine, Central South University, Changsha 410078 Powder Metallurgy Research Institute, Central South University, Changsha 410083 Department of Physics, University of Texas at Arlington, Arlington Texas 76019-0059, USA)

机构地区：[1]中南大学湘雅二医院普通外科,长沙410011 [2]中南大学湘雅医学院组织学与胚胎学教研室,长沙410078 [3]中南大学粉末冶金学院,长沙410083 [4]德克萨斯州大学物理系

出　　处：《中南大学学报（医学版）》2017年第8期874-881,共8页Journal of Central South University ：Medical Science

基　　金：湖南省发展和改革委员会课题(201583);中南大学中央高校基本科研业务费专项资金(2016zzts537)~~

摘　　要：目的:研究原卟啉(protoporphyrin IX,PpIX)介导的光动力疗法对结肠癌HCT116细胞的杀伤和促凋亡作用及其可能机制。方法:0,0.5,1,2,4,8,12μg/m L的PpIX处理后给予0,5,10 J/cm^2光照,用细胞增殖检测试剂盒(cell counting kit 8,CCK8)检测HCT116细胞存活率。用Hoechst 33342染色法及流式细胞术检测空白对照组、单光照组、单Pp IX组、光动力组细胞的凋亡情况。Western印迹检测细胞凋亡相关蛋白bcl-2,bax,caspase-3的表达。ROS试剂盒及流式细胞术检测ROS的产量变化。结果:相同的光照剂量时,细胞存活率随着PpIX剂量增大而逐渐降低(P<0.05)。相同的光敏剂浓度(<12μg/m L)时,细胞存活率随着光照剂量的增强而降低(P<0.05)。Hoechst 33342染色发现光动力组凋亡细胞相比其他3组高。流式细胞仪检测发现光动力组的HCT116细胞凋亡率、ROS产量均高于其他3组(P<0.05)。Western印迹检测发现光动力组bax,caspase-3蛋白表达增加,bcl-2蛋白表达降低(P<0.05)。结论:Pp IX介导的光动力疗法能促使HCT116细胞凋亡,ROS含量提高,可能与线粒体凋亡途径有关,通过多因素综合作用发挥效应。Objective： To explore the effects of protoporphyrin IX（Pp IX）-mediated photodynamic therapy（PDT） on induction of apoptosis and death in colon cancer cell and the underlying mechanisms. Methods： The cell killing effect of PDT on HCT116 cell was determined by cell counting kit（CCK）. The cells were divided into a control group, a single light group, a single Pp IX group, and a PDT group. Hoechst 33342 and flow cytometry was used to assess the cell apoptosis. Western blot was employed to analyze the expressions of bcl-2, bax, and caspase-3. Reactive oxygen species（ROS） was detected by flow cytometry.Results：The viability of HCT116 cell was decreased gradually with the increase of irradiation dose（P〈0.05）.Compared to the other 3 groups,ROS production,the number of apoptotic cells and the protein expressions of bax and caspase-3 in PDT group increased,while bcl-2 expression was decreased（P〈0.05）.Conclusion：Pp IX-mediated PDT can enhance the apoptosis in HCT116 cell,which may be related to mitochondrial apoptosis pathway.

关 键 词：光动力疗法 原卟啉IX 结肠癌 杀伤 凋亡 ROS 

分 类 号：R735.35[医药卫生—肿瘤]