IGF-Ⅱ对阿霉素诱导人肝癌细胞HepG2凋亡的拮抗作用研究

目的 研究胰岛素样生长因子Ⅱ(insulin-like growth factor Ⅱ,IGF-Ⅱ)对阿霉素(adriamycin,ADM)诱导的人肝癌细胞HepG2凋亡的影响及其与凋亡抑制蛋白survivin表达的关系.方法 实验分组:A组:空白对照组;B组:200 ng/ml ADM组;C组:2 n/ml IGF-Ⅱ+200 ng/ml ADM组;D组:20 ng/ml IGF-Ⅱ+200 ng/ml ADM组;E组:200 ng/ml IGF-Ⅱ+200 ng/ml ADM组,分别处理HepG2细胞48 h后,应用MTF检测各组细胞活力,流式细胞仪检测细胞的凋亡率,Western blot检测细胞中survivin蛋白的表达.结果 A、B、C、D、E组的HepG2细胞活力分别为:0.568±0.025、0.201±0.020、0.232±0.027、0.268±0.013、0.304±0.019,凋亡率分别为:6.9%±1.3%、35.4%±2.1%、31.2%±2.2%、26.4%±1.7%、21.7%±1.9%,survivin与β-actin的比值分别为:0.527±0.039、0.147±0.081、0.311±0.069、0.421±0.033、0.469±0.031,结果显示IGF-Ⅱ+ADM组的HepG2细胞活力较ADM组明显好转(P＜0.01),凋亡率显著降低(P＜0.01),IGF-Ⅱ+ADM组的HepG2细胞中survivin蛋白的表达较ADM组明显增高(P＜0.01),且IGF-Ⅱ的上述作用随IGF-Ⅱ浓度的增高而增强(P＜0.05).结论 IGF-Ⅱ能够上调HepG2细胞中survivin蛋白的表达,拮抗ADM诱导的HepG2细胞凋亡.

IGF-Ⅱ inhibits adriamycin-induced apoptosis of HepG2

Objective To investigate the effects of IGF-Ⅱ on adriamycin-induced apoptosis of hepatocellular carcinoma cell line(HepG2).Methods Cultured HepG2 cells were divided into:①group A:control group;②group B:ADM group(200 ng/ml);③group C:2 ng/ml IGF-Ⅱ+200 ns/ml ADM group;④group D:20 ng/ml IGF-Ⅱ+200 ng/ml ADM group;⑤group E:200 ng/ml IGF-Ⅱ+200 ng/ml ADM group. After HepG2 were treated for 48 h,MTT colorimetry was performed to determine the cell viability,Flow cytometry was used to detect the cell apoptosis rate,and Weastern-blot was performed to evaluate the expression of survivin. Results The cell viability of group A、B、C、D、E was respectively 0.568±0.025、0.201±0.020、0.232±0.027、0.268±0.013 and 0.304±0.019;The cell apoptosis rate of group A、B、C、D;E was respectively 6.9％±1.3％、35.4％±2.1％、31.2％±2.2％、26.4％±1.7％and 21.7％±1.9％;Survivinβ-actin of group A、B、C、D、E was respectively 0.527±0.039、0.147±0.081、0.311±0.069、0.421±0.033 and 0.469±0.031.The cell viability in IGF-Ⅱ+ADM group was better than ADM group(P＜0.01),the cell apoptosis rate in IGF-Ⅱ+ADM group was significantly lower than ADM group(P＜0.01),the expression level of survivin in IGF-Ⅱ +ADM group was significantly higher than ADM group(P＜0.01). Conclusions IGF-Ⅱ inhibits ADM-induced apoptosis of HepG2 cells,probably by up-regulating cell's survivin expression.