摘要
目的体外实验探讨MEK1特异性抑制剂PD98059对奥沙利铂药物治疗作用的影响及PUMA在这一过程中的作用。方法MTT法检测转染MEK1active质粒后及不同药物处理后对LoVo细胞增殖率的影响;Western blot检测奥沙利铂和/或PD98059处理后PUMA蛋白的表达和ERK的活性;利用表达反义PUMA基因的细胞克隆抑制PUMA的表达后检测对PD98059和奥沙利铂诱导凋亡作用的影响。结果在肠癌LoVo细胞中,ERK的激活增加细胞的增殖率;奥沙利铂可以抑制ERK的活性,并呈剂量依赖关系;PD98059可以协同奥沙利铂抑制细胞增殖率的作用;奥沙利铂和PD98059可以协同诱导PUMA的表达;抑制PUMA的表达可以减弱奥沙利铂和PD98059诱导的LoVo细胞的凋亡。结论PD98059通过诱导PUMA表达增强肠癌LoVo细胞对奥沙利铂的敏感性。
Objective To investigate the effects of MEK1 specific inhibitor PD98059 on oxaliplatin-treated colorectal cancer cells and the potential mechanism. Methods Cell proliferation was assessed by MTY assay after transfecting MEK1 active plasmid into LoVo cells. LoVo cells were treated with oxaliplatin or PD98059, and the proliferation was assessed by MTY assay. PUMA expression and ERK activity were determined by Western blot. Apoptosis was assessed by Hoechst 33258 dye after PUMA expression was suppressed. Results Increasing activity of ERK enhanced the proliferation of LoVo cells. The activity of ERK was suppressed by oxaliplatin. PD98059 and oxaliplatin decreased the proliferation rate of LoVo cells synergistically. PUMA expression increased after PD98059 and oxaliplatin treatment. The suppression of PUMA expression by stably transfecting PUMA anti-sense vector decreased apoptosis induced by oxaliplatin and PD98059. Conclusion PD98059 enhances the effects of oxaliplatin on colorectal cancer cells mediated by PUMA expression.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2007年第17期1696-1698,共3页
Journal of Third Military Medical University
