IGF-Ⅱ对阿霉素诱导人肝癌细胞HepG2凋亡的拮抗作用研究

IGF-Ⅱ inhibits adriamycin-induced apoptosis of HepG2

目的研究胰岛素样生长因子Ⅱ（insulin-like growth factorⅡ，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：200ng／ml ADM组；C组：2ng／ml IGF-Ⅱ＋200ng／ml ADM组；D组：20ng／ml IGF-Ⅱ＋200ng／ml ADM组；E组：200ng／ml IGF-Ⅱ＋200ng／ml ADM组，分别处理HepG2细胞48h后，应用MTT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。

Objective To investigate the effects of IGF-Ⅱ on adriamycin-induced apoptosis of hepatocenular carcinoma cell line（HepG2）. Methods Cultured HepG2 cells were divided into： 1.group A： control group; 2.group B： ADM group（200 ng/ml） ; 3.group C： 2 ng/ml IGF-Ⅱ ＋ 200 ng/ml ADM group; 4.group D：20 ng/ml IGF-Ⅱ ＋200 ng/ml ADM group; 5.group 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% ; Survivin/β-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.