抑制核转录因子κB活性对SGC7901/VCR细胞mdr1表达及凋亡的影响  

Effects of inhibition of nuclear factor-κB activity on mdr1 expression and apoptosis of SGC7901/ VCR cells

作　　者：陈庆[1] 陈虎[1] 郑海[1] 陈道达[1] 蒋春舫[1] 周洁[2] 

机构地区：[1]华中科技大学同济医学院附属协和医院外科,武汉430022 [2]华中科技大学同济医学院基础医学院生物化学与分子生物学教研室

出　　处：《中华实验外科杂志》2010年第8期1066-1068,共3页Chinese Journal of Experimental Surgery

基　　金：湖北省自然科学基金资助项目（2007ABA065）;湖北省卫生厅科技攻关计划资助项目（JX1B006）

摘　　要：目的 观察突变型IκBα抑制核转录因子κB（NF-κB）活性对耐药胃癌细胞株SGC7901/VCR中多药耐药基因1（mdr1）的表达及细胞凋亡的影响.方法 突变型IκBα真核表达重组体（pCMV4-mIκBα）经脂质体介导转染至SGC7901/VCR细胞中,凝胶迁移率实验（EMSA）检测SGC7901/VCR细胞核内NF-κB活性,逆转录-聚合酶链反应（RT-PCR）和Western blot检测细胞内mdr1的表达,荧光分光光度法检测罗丹明123（Rh123）的胞内聚集,流式细胞术检测细胞凋亡.结果 pCMV4-mIκBα瞬时转染人SGC7901/VCR细胞24 h后,可显著抑制NF-κB活性达64%,抑制mdr1的表达达75%,使Rh123在细胞内的聚集增加2.3倍.转染24 h后SGC7901/VCR细胞凋亡率为（14.15±1.52）,与对照组（4.37±1.31）比较差异有统计学意义（P＜0.01）.结论 突变型IκBα抑制NF-κB活性对耐药胃癌细胞株mdr1的表达有直接抑制作用,并促使耐药胃癌细胞凋亡.Objective To investigate the effects of inhibition of nuclear factor-κB （NF-κB） activity induced by mutated IκBα on mdrl expression and apoptosis of gastric carcinoma cell lines SGC7901/ VCR. Methods pCMV4-mκBα was transfected into SGC7901/VCR cells by lipofectamine 2000.The activity of NF-κB was detected by EMSA. Reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting were used to detect the mdrl gene expression. The accumulation of Rhodamine123 （Rh123） in SGC7901/VCR cells was analyzed by fluorescence spectrophotometric method. The apoptosis of SGC7901/VCR cells was examined by FCS technique. Results The inhibition rate of NF-kB activity and mdrl expression in SGC7901/VCR cells transfected with pCMV4-mlκBα was 64% and 75% respectively as compared with the control cells. Twenty-four h after transfection of mlκBα plasmid, the intracellular Rhl23 concentration was increased by 2.3-fold. The apoptosis rate in mlκBα plasmid transfected goups （14.15 ±1.52） was increased significantly as compared with the control group （P〈0.01）. Conclusion Inhibition of NF-κB activity results in the decrease of mdrl gene expression and increase of apoptosis by inhibiting drugs efflux in SGC 7901/VCR cells.

关 键 词：胃癌 核因子-ΚB 多药耐药基因1 脱噬作用 

分 类 号：R735.2[医药卫生—肿瘤]