二氯乙酸联合顺铂对人结肠癌HCT116细胞凋亡的影响  

The influence of dichloroacetate combined with cisplatin on the apoptosis of colorectal carcinoma cells HCT116

作　　者：孙洁[1] 孟祥军[1] 

机构地区：[1]上海交通大学附属第一人民医院消化内科,200080

出　　处：《中华普通外科杂志》2015年第2期138-141,共4页Chinese Journal of General Surgery

基　　金：国家自然科学基金资助项目（81272745）

摘　　要：目的 探讨二氯乙酸（dichloroacetate,DCA）联合顺铂对人结肠癌HCT116细胞凋亡的影响及其可能的机制.方法 单独或联合使用DCA和顺铂处理HCT116细胞,检测其对细胞增殖及凋亡的影响;用罗丹明123染色细胞,在荧光显微镜下观察线粒体跨膜电位变化;定量-PCR检测bcl-2的表达情况;检测HCT116细胞中半胱氨酸天门冬氨酸蛋白酶3（cysteinyl aspartate specific proteinase-3,caspase-3）蛋白的活性.结果 单用DCA或顺铂对HCT116细胞增殖均有抑制作用,且呈时间-剂量依赖性.与同浓度单药相比,联合应用DCA和顺铂处理细胞48 h后对细胞凋亡出现显著的协同效应.联合用药组的细胞核浓聚或碎裂、线粒体跨膜电位降低更明显,且下调bcl-2表达的幅度与单药组比较差异均有统计学意义（均P ＜0.05）.联合用药组caspase-3活性高于单药组,差异有统计学意义（P＜0.01）.结论 DCA对人结肠癌HCT116细胞有增殖抑制和促凋亡作用,具有一定的时间-剂量依赖性.DCA联合顺铂对结肠癌HCT116细胞增殖和凋亡作用的影响显著增加,可能与线粒体跨膜电位降低和bcl-2的表达下调等有关.Objective To explore the effects of dichloroacetate （DCA） combined with cisplatin on the apoptosis of HCT116 and possible mechanisms.Methods The inhibitory effects of DCA and cisplatin alone or in combination on colorectal carcinoma cell line HCT116 were examined by MTT and Hoechst 33342 staining,the mitochondrial membrane potential changes were measured by Rodanmine123 staining under fluorescent microscope.The expression of bcl-2 was checked by qPCR.The activity of caspase-3 was assayed.Results DCA or cisplatin alone could inhibit the growth of HCT116 in a time and dose dependent manner.Compared with single drug treatment,there was significantly synergistic effect after treatment of DCA combined with cisplatin for 48 hours.Compared with the single drug treatment,the nuclear morphological changes such as chromatin condensation and fragmentation were more severe,and the mitochondrial transmembrane potential declines were markedly apparent for DCA ＋ cisplatin group.The expression of bcl-2 gene in combination group was inhibited （P 〈 0.05）,and the activity of caspase-3 significantly increased （P 〈 0.01）.Conclusions DCA could inhibit the proliferation and induce the apoptosis of HCT116 cells in a time and dose dependent manner.The combination use of DCA and cisplatin has a synergistic effect on the biological action of HCT116.This may be attributed to lowering of mitochondrial transmembrane potential and the suppressed expression of bcl-2 gene.

关 键 词：结肠肿瘤 二氯乙酸盐 顺铂 细胞凋亡 

分 类 号：R971.1[医药卫生—药品]