阿霉素联合p21^(CIP1)基因转染对人肝母细胞瘤细胞株HepG2增殖的影响  

Inhibitory Effect of Adriamycin Combined P21^(CIP1) Gene Transfection on Proliferation of Human Hepatoblastoma Cell Line HepG2

作　　者：熊娟[1] 胡丽华[1] 李一荣[1] 王琳[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院免疫学研究所,湖北武汉430022

出　　处：《癌症》2008年第5期476-481,共6页Chinese Journal of Cancer

基　　金：教育部高等学校博士学科点专项科研基金资助项目(No.20060487045)~~

摘　　要：背景与目的:阿霉素类药物是治疗肝母细胞瘤的传统化疗药,但近年来其疗效不佳是困扰临床的一大问题,目前配合基因治疗提高阿霉素的疗效,是肝母细胞瘤治疗发展的新趋势。本实验研究阿霉素与p21基因转染联合对人肝母细胞瘤HepG2细胞增殖的影响。方法:实验分为空白对照组、单独加药组、pcDNA3转染对照组、p21转染组及p21转染+药物联合组。用四甲基偶氮唑蓝(MTT)法检测转染后HepG2细胞的生长趋势,联合处理后细胞增殖抑制状况;荧光定量PCR检测转染后p21mRNA的表达情况,联合处理后survivinmRNA的水平变化。结果:转染后,p21转染组在第3d和第4d的生长速度明显慢于两对照组(P<0.01);经鉴定其有p21mRNA表达量的增高,是空白对照组的155倍(P<0.05)。以空白组为对照,在第3～5d,联合组增殖抑制率明显高于单独加药组和p21转染组(第3d:43.92%vs.32.97%、35.77%,P<0.01;第4d:59.86%vs.39.35%、40.96%,P<0.01;第5d:51.81%vs.33.91%、10.68%,P<0.01);且1～4d内随联合用药时间的延长,抑制效应增强(r=0.91,P<0.05),并在第4d时较显著(Q=1.07)。荧光定量PCR显示,联合组survivinmRNA水平显著低于p21转染组(P<0.01);与单独加药组比较,联合作用仅在48h时差异有统计学意义(P<0.01)。结论:在一定时间范围内p21可增强阿霉素对HepG2细胞的增殖抑制作用,降低胞内survivinmRNA的表达。BACKGROUND ＆ OBJECTIVE： Chemotherapy protocols using adriamycin （ADM） is a standard treatment for hepatoblastoma, but the treatment results became unsatisfied because of drug resistance. Recently, ADM combined gene therapy is a developing alternative treatment for hepatoblastoma. This study was to investigate the effect of ADM combined human p21^CIP1 transfection on the proliferation of hepatoblastoma cell line HepG2. METHODS. HepG2 cells were divided into empty control group （no treatment）, ADM group （treated with 0.5 μg/mL ADM）, blank control group （transfected with blank plasmid pcDNA3）, p21 group （transfected with plasmid pcDNA3-p21）, and combination group （ADM treatment plus p21 transfection）. The proliferation of HepG2 cells was observed by M-IF assay. The mRNA levels of p21 and survivin were detected by real-time polymerase chain reaction （PCR）. RESULTS： After transfection, the mRNA level of p21 in p21 group was increased by 155 folds of that in empty control group （P〈 0.05）. p21 inhibited the proliferation of HepG2 cells at Day 3 and Day 4 after transfection （P〈0.01）. The proliferation inhibition rate was significantly higher in combination group than in ADM group and p21 group （43.92% vs. 32.97% and 35.77% at Day 3, P〈0.01; 59.86% vs. 39.35% and 40.96% at Day 4, P〈0.01; 51.81% vs. 33,91% and 10.68% at Day 5, P〈0.01）. This effect was enhanced along with the increasing time of co-treatment from Day 1 to Day 4 （r=0.91, P〈0.05）, and it was obvious at Day 4 （Q =1.07）, The mRNA level of survivin was significantly lower in combination group than in p21 group and ADM group （P〈0.01）, CONCLUSION： p21 gene transfection plus ADM can inhibit the proliferation of HepG2 cells and down-regulate the level of survivin mRNA, thus may be a potential therapeutic strategy against human hepatoblastoma.

关 键 词：肝脏肿瘤 HepG2细胞 阿霉素 p21^CIP1基因 增殖 

分 类 号：R735.7[医药卫生—肿瘤]