咖啡因在骨肉瘤细胞株热化疗中增效作用的实验研究

Enhancement effect of caffeine in the combined treatment of osteosarcoma OS－ 732 cell line with hyperthermia and cisplatin

目的探讨咖啡因结合顺铂热化疗对骨肉瘤细胞株 (OS－ 732)细胞毒性的影响。方法 43℃恒温条件下，骨肉瘤细胞株 (OS－ 732)分别经咖啡因、顺铂、咖啡因＋顺铂作用 1 h，分别采用体外 3－ (4， 5－ dimethylthiazol－ 2－ yl)－ 2,5－ diphenyltetrazolium bromide(MTT法 )细胞毒性试验和流式细胞仪 (FCM)分析，观察三者对细胞毒性及细胞周期的影响。结果咖啡因浓度为 0、 0.2、 2.0、 5.0、 10.0、 20.0 mmol/L时，其细胞毒性指数分别为 15.79％、 19.74％、 19.74％、 21.05％、 38.16％、 46.05％。顺铂浓度为 0、 0.1、 1.0、 10.0μ g/ml时，其细胞毒性指数分别为 15.79％、 25.00％、 30.26％、 44.74％。而浓度为 2.0、 5.0 mmol/L的咖啡因与浓度为 0.1、 1.0、 10.0μ g/ml的顺铂联合作用，其细胞毒性指数明显增加，最高达 64.47％。且结合 FCM分析显示其对细胞周期也有明显的影响， G0与 G1期细胞比例明显增加， G2与 M期明显减少。结论安全浓度的咖啡因在骨肉瘤细胞株 (OS－ 732)顺铂热化疗中有明显的增效作用。

Objective To observe the cytotoxicity effect of caffeine combined with hyperthermia and cisplatin on osteosarcoma OS－ 732 cell line. Methods At 43℃ , the osteosarcoma OS－ 732 cell line was treated with caffeine, cisplatin, caffein＋ cisplatin for 1 hour, respectively, by using 3－ (4， 5－ dimethylthiazol－ 2－ yl)－ 2, 5－ diphenyltetrazolium bromide(MTT) assay in vitro. The cytotoxicity effect was observed with flow cytometry (FCM),and the cell cycle phases were analysed. Results At 43℃ , the osteosarcoma OS－ 732 cell line was treated with caffeine(0、 0.2、 2.0、 5.0、 10.0、 20.0 mmol/L), the cytotoxicity index were 15.79％、 19.74％、 19.74％、 21.05％、 38.16％、 46.05％ , respectively; with cisplatin treatment (0、 0.1、 1.0、 10.0μ g/ml), the cytotoxicity index were 15.79％、 25.00％、 30.26％、 44.74％ respectively. Caffeine and cisplatin treatment used simultaneously, the cytotoxicity index increased obviously. The cell cycle of the treated OS－ 732 cell line was changed with marked increase in the G0/G1 phase and decrease in G2/M phase. Conclusion Nontoxic dose of caffeine combined with hyperthemia and cisplatin had enhancement of the cytotoxicity effect on osteosarcoma OS－ 732 cell line.