抑制PDK对人结肠癌细胞增殖和化疗敏感性影响及其机制的探讨  

Effect and mechanism of PDK inhibition on cell proliferation and chemotherapy in human colon cancer cell lines

作　　者：童晶涛[1] 谢赣丰[1] 欧娟娟[1] 萨日娜[1] 江恒[1] 梁后杰[1] 

机构地区：[1]第三军医大学第一附属医院肿瘤科,重庆400038

出　　处：《中华肿瘤防治杂志》2010年第9期644-647,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30873015)

摘　　要：目的:探讨抑制丙酮酸脱氢酶激酶(PDK)对结肠癌细胞增殖和化疗敏感性的影响及可能机制。方法:应用MTT法观察PDK抑制剂(DCA)对人结肠癌细胞化疗增敏效应;应用Hoechst33258染色通过激光共聚焦显微镜观察DCA诱导的LoVo细胞凋亡;用流式细胞术观察DCA诱导的细胞凋亡、线粒体膜电位和Ca2+通道开放的情况。结果:抑制PDK(DCA)能明显影响结肠癌细胞增殖,其中90mmol/LDCA刺激SW620、LoVo、HT29、LS174T和293T细胞48h后,活性细胞率分别为(23.90±2.79)%、(5.90±1.31)%、(32.82±4.50)%、(29.72±3.03)%和(84.86±0.50)%。与小剂量DCA(2.5和5mmol/L)联合作用LoVo细胞48h,低剂量5-FU(5mmol/L)的活性细胞率由(66.03±3.95)%分别降至(50.78±2.05)%和(39.8±2.86)%;而低剂量奥沙利铂(0.5mmol/L)的活性细胞率由(71.18±0.62)%,分别降至(58.24±3.27)%和(53.17±2.65)%。30mmol/LDCA作用4种结肠癌细胞48h后,LS174T、SW620、HT29、LoVo和293T凋亡的发生率分别为(22.27±1.75)%、(14.42±2.36)%、(25.55±0.78)%、(13.20±1.58)%和(10.73±1.49)%。30mmol/LDCA作用LoVo细胞48h后,线粒体膜电位值由4.04±1.47下降为2.09±0.86;胞内Ca2+浓度下降了83.6%。结论:抑制PDK能抑制结肠癌细胞增殖活性,增加化疗敏感性。该作用可能是通过线粒体功能异常引起结肠癌细胞凋亡。OBJECTIVE：To study the role and mechanism of PDK（pyruvate dehydrogenase kinase）inhibition on cell proliferation and chemotherapy in human colon cancer cell lines.METHODS：MTT assay was used to analyse cell growth;Hoechst33258 staining was used to describe apoptosis;apoptosis,mitochondria membrane voltage and Ca2＋ channel were all analyzed by flow cytometry.RESULTS：DCA inhibited cellular proliferation.The 90 mmol/L DCA stimulated SW620,LoVo,HT29,LS174T and 293T for 48 h,then the rate of cell viability was（23.90±2.79）%,（5.90±1.31）%,（32.82±4.50）%,（29.72±3.03）% and（84.86±0.50）%.Treated LoVo by 5-FU（5 μmol/L）combined with DCA（2.5 and 5 mmol/L）for 48 h,the rate of cell viability was from（66.03±3.95）% to（50.78±2.05）% and（39.8±2.86）%;Treated LoVo by L-OHP（5 μmol/L）combined with DCA（2.5 and 5 mmol/L）for 48 h,the rate of cell viability was from（71.18±0.62）% to（58.24±3.27）% and（53.17±2.65）%.Forty-eight hours after co-cultured with 30mmol/L DCA,the rate of SW620,LoVo,HT29,LS174T,293T cells apoptosis was（22.27±1.75）%,（14.42±2.36）%,（25.55±0.78）%,（13.20±1.58）% and（10.73±1.49）%.When LoVo was treated with 30 mmol/L DCA for 48 h,the value of mitochondrial transmembrane electrical potential was from 4.04±1.47 to 2.09±0.86;and the decrease rate of intracellular calcium levels was 83.6%.CONCLUSION：Iinhibiting PDK may stimulate anticancer activity via inducing cellular apoptosis,changing with the mitochondria membrane voltage and Ca2＋ channel.

关 键 词：结肠肿瘤/病理学 结肠肿瘤/药物疗法 细胞增殖 细胞凋亡 

分 类 号：R735.35[医药卫生—肿瘤]