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目的:探讨mTOR抑制剂雷帕霉素与顺铂联合应用对宫颈癌Hela细胞生长的抑制作用及对HIF-1α和VEGF表达的影响。方法:分别以雷帕霉素、顺铂及雷帕霉素联合顺铂处理体外培养人宫颈癌Hela细胞,采用MTT比色法检测人宫颈癌Hela细胞的抑制率,金氏公式评价两药联合用药的效果,采用RT-PCR法、蛋白质印迹法检测各组细胞HIF-1α和VEGF mRNA和蛋白的表达。结果:雷帕霉素和顺铂对宫颈癌Hela细胞的生长有明显的抑制作用,并呈剂量依赖性,分别联合应用2种浓度雷帕霉素(10和20nmol/mL)与顺铂(0.25和0.5mg/mL)协同治疗指数q值均>1.15,两者有协同作用。雷帕霉素与顺铂联合用药HIF-1αmRNA的表达为0.242±0.048,单独用药雷帕霉素组为0.428±0.068,顺铂组为0.357±0.051;雷帕霉素与顺铂联合用药组VEGF mRNA的表达为0.498±0.093,单独用药雷帕霉素组为0.651±0.112,顺铂组为0.623±0.125,联合用药组与单独用药组比较表达量明显降低,P<0.05;雷帕霉素与顺铂联合用药组HIF-1α蛋白的表达为0.514±0.092,单独用药雷帕霉素组为0.625±0.132,顺铂组为0.635±0.120;雷帕霉素与顺铂联合用药组VEGF蛋白的表达为0.409±0.082,单独用药雷帕霉素组为0.650±0.114,顺铂组为0.623±0.102,联合用药组与单独用药组比较表达量明显降低,P<0.05。结论:单独及联合应用雷帕霉素与顺铂对宫颈癌Hela细胞的生长均有抑制作用,两药联合应用有明显的协同作用,雷帕霉素联合顺铂具有明显下调HIF-1α、VEGF基因和蛋白表达的作用。
Objective: To investigate the inhibitory effect of mTOR inhibitor rapamycin and cisplatin on the growth of cervical cancer Hela cells and its effect on the expression of HIF-1α and VEGF. Methods: Human cervical cancer Hela cells were treated with rapamycin, cisplatin, rapamycin and cisplatin respectively. MTT assay was used to detect the inhibitory rate of Hela cells. King’s formula was used to evaluate the efficacy of the two drugs The effects of the drugs, the expression of HIF-1α and VEGF mRNA and protein in each group were detected by RT-PCR and Western blot. Results: Rapamycin and cisplatin significantly inhibited the growth of cervical cancer Hela cells in a dose-dependent manner. The combination of two concentrations of rapamycin (10 and 20 nmol / mL) and cisplatin (0.25 And 0.5mg / mL) synergistic treatment index q values were> 1.15, the two have a synergistic effect. The expression of HIF-1αmRNA in rapamycin and cisplatin group was 0.242 ± 0.048, the group of rapamycin alone was 0.428 ± 0.068, the group of cisplatin was 0.357 ± 0.051; The combination of rapamycin and cisplatin group VEGF mRNA expression was 0.498 ± 0.093, the single drug rapamycin group was 0.651 ± 0.112, cisplatin group was 0.623 ± 0.125, the combination group and the drug alone group was significantly lower expression, P <0.05; rapamycin and The expression of HIF-1α protein was 0.514 ± 0.092 in cisplatin combination group, 0.625 ± 0.132 in rapamycin alone group and 0.635 ± 0.120 in cisplatin group. The VEGF protein expression in rapamycin and cisplatin group was significantly higher than that in cisplatin group 0.409 ± 0.082, 0.650 ± 0.114 in the rapamycin group alone and 0.623 ± 0.102 in the cisplatin group. The expression level of the combination group was significantly lower than that of the drug alone group (P <0.05). Conclusion: Rapamycin and cisplatin alone and in combination have inhibitory effect on the growth of cervical cancer Hela cells. The combination of the two drugs has obvious synergistic effect. Rapamycin and cisplatin can significantly reduce the expression of HIF-1α, VEGF The role of genes and protein expression.