DENND2D通过调控PARP1对非小细胞肺癌细胞系H1299中顺铂细胞毒性的影响  

Effect of DENND2D on the cisplatinum cytotoxicity in H1299 cells by regulating PARP1

作　　者：凌兵[1] 冯林[1] 程书钧[1] 

机构地区：[1]中国医学科学院北京协和医学院肿瘤医院肿瘤研究所病因与癌变实验室,北京100021

出　　处：《癌变．畸变．突变》2013年第2期87-90,95,共5页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：973计划资助项目(2004CB518707)

摘　　要：目的:检测肺癌细胞系H1299细胞中DENN/MADD domain containing2D(DENND2D)基因过表达对顺铂细胞毒性的影响,并初步探讨其机制。方法:应用瞬时和稳定转染两种方法使H1299细胞外源过表达DENND2D基因,以空载体组作为对照组,应用CCK-8比色法检测不同浓度顺铂作用下H1299细胞的存活情况,据此计算出IC值。利用Western blot方法检测外源过表达50DENND2D的H1299细胞多聚ADP核糖聚合酶[poly(ADP-ribose)polymerase-1,PARP1]的表达情况。利用顺铂处理外源过表达DENND2D的H1299细胞,并检测不同处理时间点(0、1、2、4和8h)H1299细胞多聚ADP核糖[poly(ADP-ribose),PAR]的表达情况。结果:顺铂对外源过表达DENND2D基因的H1299细胞毒性明显高于空载体组(IC值降低,P<0.05)。外源过表达DENND2D的50H1299细胞PARP1蛋白和PAR蛋白的表达均比空载体对照组低,且空载体对照组PAR的表达随时间推移呈上升趋势,而外源过表达DENND2D组则呈下降趋势。结论:DENND2D可能通过对PARP1的调控增强了顺铂对肺癌细胞系H1299的细胞毒性。OBJECTIVE： To investigate the effect of DENN/MADD domain containing 2D （DENND2D） gene on the cytotoxity of cisplatinum in non-small cell lung cancer cell line H1299 and explore the mechanism preliminarily. METHODS：DENND2D gene was transient stably transfected into H1299 cells. CCK-8 assay was used to test the cell counts of DENND2D transfected H1299 cells treated with various concentrations of cisplatinum and IC50 results were attained. PARP1 protein expression was evaluated by western blot. DENND2D over-expressed H1299 cells were treated with cisplatinum for 0,1,2,4 and 8h. The PAR expressions were measured by Western blot. RESULTS：IC50 was decreased in DENND2D over-expressed group compared to vector group. PARP1 and PAR were down-regulated in DENND2D over-expressed H1299 cells. With the prolonged platinum treatment time,the expression of PAR was down-regulated in DENND2D group whereas it was opposite in the vector group. CONCLUSION：DENND2D may enhanced the cytotoxicity of cisplatinum in H1299 cells by down regulating PARP1.

关 键 词：DENND2D PARP1 顺铂 非小细胞肺癌 细胞毒性 

分 类 号：R730.5[医药卫生—肿瘤]