胰岛素抵抗肝癌细胞IGF-1R/NF-κB表达及多药耐药机制研究

目的 探讨胰岛素抵抗（IR）肝癌细胞胰岛素样生长因子1受体（IGF-1R）和核因子-κB（NF-κB）表达变化及多药耐药（MDR）发生机制。方法 采用高浓度胰岛素诱导人肝癌细胞（HepG2和HepG2.2.15）建立胰岛素抵抗（IR）细胞模型。采用Western blot 法检测胰岛素受体（InsR）、IGF-1R、NF-κB 和 P-糖蛋白（P-gp）表达变化。使用流式细胞仪（Annexin V-FITC法）检测阿霉素对细胞凋亡的影响。结果 分别用100 nmol/L 和 1 000 nmol/L 胰岛素培养 HepG2 和 HepG2.2.15 细胞 48 h，成功建立 IR 肝癌细胞模型；IR 肝癌细胞 IGF-1R、NF-κB、P-gp 表达上调，而InsR 表达下调；应用 25μg/mL 阿霉素作用细胞 24 h 后，IR-HepG2 细胞组凋亡率（31.1%±1.9%）显著低于HepG2 细胞组【（49.7%±2.2%），P＜0.01】，IR-HepG2.2.15细胞凋亡率【（20.1±1.7） %】显著低于 HepG2.2.15 细胞【（33.8±1.8）%，P＜0.01】；HepG2.2.15 和 IR-HepG2.2.15 细胞凋亡率分别较 HepG2 和 IR-HepG2 细胞显著降低（P＜0.01）。结论 IGF-1R/NF-κB/P-gp 过表达可能介导 IR 肝癌细胞对阿霉素的多药耐药。

Expression of IGF-1R/NF-kappa B and multi-drug resistance in insulin resistance-HepG2 and HepG2.2.15 cells in vitro

Objective To investigate the expression of insulin-like growth factor 1 receptor（IGF-1R），nuclear factor-κB（NF-κB） and the mechanism of multi-drug resistance（MDR） in insulin resistance（IR）- hepatoma cells in vitro. Methods The human hepatoma cells（HepG2 and HepG2.2.15） were induced by high concentration of insulin to establish a insulin resistance model. The expression of insulin receptor （InsR），IGF-1R，NF kappa B，P glycoprotein（P-gp） were detected by Western bloting and the effect of adriamycin on cell apoptosis were detected by flow cytometry（annexin V-FITC）. Results IR model of hepatoma cells were established successfully by subjecting the HepG2 and HepG2.2.15 cells to 100 nmol/L and 1 000 n mol/L insulin respectively for 48 hours. The expression of IGF-1R，NF-kappa B and P-gp were up-regulated in IR hepatoma cells，while the InsR expression was down-regulated；After treatment of the two cells with 25 μg/ml doxorubicin for 24 hours，the apoptosis rate of IR-HpeG 2 cells （31.1% ±1.9%） was significantly lower than HepG2 cells （49.7±2.2）%，P＜0.01]，and the apoptosis rate of IR-HepG2.2.15 cells（20.1±1.7）% was significantly lower than HepG2.2.15 cells（33.8±1.8）%，P＜0.01]；The apoptotic rates of HepG2.2.15 or IR-HepG2.2.15 cells were significantly lower than HepG2 or IR-HepG2 cells（P＜0.01），respectively. Conclusion The expression of IGF-1R， NF-kappa B and P-gp are up-regulated in IR-hepatoma cells，which might induce the MDR to adriamycin.