番茄红素对人骨肉瘤细胞顺铂增敏作用及机制研究

摘要

Objective To investigate the efficacy enhancing effect and mechanism of lycopene (LP) on human steosarcoma MG-63 cells treated by Cisplatin.Methods The human osteosarcoma MG-63 cells in logarithmic growth phase was treated with LP (l0μg/ ml),Cisplatin (40μg/ml) and LP (10μg/ml) + Cisplatin (20μg/ml),and blank group was set.The cellular growth inhibition rate was calculated by MTT.Cell cycle and apoptosis rate were analyzed by flow cytometry(FCM).The expression of Bax mRNA and bcl-2 mRNA were detected by RT-PCR and the ratio of Bax/bcl-2 was calculated.The expression of Caspase-3 protein was detected by western blotting.Results Compared with Cisplatin group,the cellular growth inhibition rate of human osteosarcoma M G-63 cell in LP + Cisplatin group was significantly increased (P ＜ 0.05).The cell cycle was arrested at G0/G1 phase(P ＜ 0.05).The apoptosis rate was significantly increased(P ＜ 0.01).The expression of Bax mRNA was increased,while the expression of bcl-2 mRNA was significantly decreased (P ＜ 0.05) and the ratio of Bax/bcl-2 was significantly increased (P ＜ 0.01).The expression of Caspase-3 protein was significantly increased(P ＜ 0.01).Gomparcd with blank group group,the G0/G1 phase in cell cycle of LP group was increased and the G2/M phase was decreased,and the expression of bcl-2 mRNA was significantly decreased (P ＜ 0.05).The ratio of Bax/bcl-2 was significantly increased (P＜0.05).The expression of Caspase-3 protein was significantly increased (P ＜0.01).Conclusion LP can effectively enhance the effect on human liver cancer MG-63 cells treated by Cisplatin,which perhaps was related to its effects of altering the cell cycle and the expression of apoptosis-related genes and proteins.%目的 研究番茄红素(lycopene,LP)对人骨肉瘤MG-63细胞顺铂化疗敏感度的影响并初步探讨其可能的作用机制.方法 体外培养并取对数生长期人骨肉瘤MG-63细胞,设空白对照组、LP组(10μg/ml)、顺铂组(40 μg/ml)和联合组[LP(10μg/ml)+顺铂(20μg/ml)];药物干预48h后,采用噻唑蓝(MTT)比色法测定细胞增殖抑制率,流式细胞术(flow cytometry,FCM)检测细胞周期和细胞凋亡状况,反转录PCR(RT-PCR)法检测Bax mRNA、bcl-2 mRNA表达并计算Bax/bcl-2比值,蛋白免疫印迹(Western blot)法检测caspase-3蛋白表达.结果 联合干预组与顺铂组比较发现,联合干预组人骨肉瘤MG-63细胞增殖抑制率显著升高(P＜0.05),细胞周期G0/G1期显著延长(P＜0.05)、S期和G2/M期显著缩短(P＜0.05,P＜0.01),细胞凋亡率(apoptosis index,AI)显著升高(P＜0.01),Bax mRNA表达明显上调、bcl-2 mRNA表达显著下调(P＜0.05)且Bax/bcl-2比值显著升高(P＜0.01),caspase-3蛋白表达上调(P＜0.01).LP组与空白对照组比较发现,LP组MG-63细胞周期明显改变(G0/G1期延长、G2/M期缩短),bcl-2 mRNA显表达著下调(P＜0.05)、Bax/bcl-2比值显著升高(P＜0.05),caspase-3蛋白表达显著上调(P＜0.01).结论 LP具有提高人骨肉瘤MG-63细胞顺铂敏感度的药理学作用,其机制可能与LP能够阻滞细胞周期以及调节凋亡相关调控基因和蛋白表达有关.