结肠癌HT-29细胞中PIDD蛋白的表达及其对细胞耐药性的影响  

The expression of PIDD in colon carcinoma HT-29 cells and its effect on drug resistance

作　　者：彭晶晶[1,2] 李建军[1] 梁后杰[1] 边志衡[1] 江恒[1] 

机构地区：[1]第三军医大学西南医院肿瘤科,重庆400038 [2]成都军区总医院肿瘤科,成都600038

出　　处：《肿瘤》2009年第4期341-344,共4页Tumor

基　　金：重庆市自然科学基金资助项目(编号:CSTC2006BB5060)

摘　　要：目的:以结肠癌HT-29细胞株为载体,研究PIDD(p53-induced protein with a death domain)蛋白在结肠癌耐药中的作用机制。方法:构建并转染PIDD基因的小干扰RNA(small interfering RNA,siRNA)片段,用Western印迹法检测其干扰效果。通过MTT比色法检测细胞对化疗药物的敏感性,并用凝胶阻滞分析(electrophoretic mobility shift assay,EMSA)法检测核因子NF-κB活性的变化。结果:RNA干扰后给予细胞药物刺激,细胞核中PIDD表达下降,核因子NF-κB活性明显降低,细胞耐药性下降。结论:转染siRNA后,HT-29细胞对5-氟尿嘧啶(5-fluorouracil,5-FU)的敏感性增加,此作用可能与细胞核PIDD复合体减少、核因子NF-κB活性减低及细胞凋亡增多有关。Objective：To study the action mechanism for the effect of p53-induced protein with a death domain （PIDD） on drug resistance of colon carcinoma HT-29 cells.Methods：PIDD small interfering RNA （siRNA） fragment was constructed and transfected it into HT-29 ceils with liposome 2000. Western blotting was used to detect the effect of interference. MTT assay was used to detect the sensitivity of HT-29 cells to chemotherapeutics. The alteration of the activity of NF-κB was tested by electrophoretic mobility shift assay （EMSA）. Results ： The expression of PIDD was decreased in HT-29 cells after RNA interference. The activity of NF-κB was significantly decreased and the drug resistance of HT-29 cells to 5-FU was decreased obviously after interfering PIDD. Conclusion：The sensitivity of HT-29 cells to 5-FU is elevated obviously after transfection of the PIDD siRNA. The effect is related with the decrease in the compound of PIDD, depression of the activity of NF-κB, and increase in cell apoptosis.

关 键 词：结肠肿瘤 细胞凋亡 抗药性 肿瘤 RNA干扰 NF—κB 

分 类 号：R735.35[医药卫生—肿瘤] R73-36[医药卫生—临床医学]