阿霉素联合p21CIP1基因转染对人肝母细胞瘤细胞株HepG2增殖的影响

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。

Inhibitory effect of adriamycin combined P21CIP1 gene transfection on proliferation of human hepatoblastoma cell line HepG2

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 p21CIP1 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 microg/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 MTT 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.