雷帕霉素对人骨肉瘤细胞增殖和凋亡的影响

Effects of rapamycin on proliferation and apoptosis of human osteosarcoma cells in vitro

目的观察雷帕霉素（RAPA）对人骨肉瘤细胞U-20S和Saos-2增殖和凋亡的影响并探讨其机制。方法体外培养骨肉瘤细胞株U-2OS和Saos-2；噻唑蓝（MTT）比色法和流式细胞仪检测不同浓度雷帕霉素对骨肉瘤细胞增殖和凋亡的影响；Westernblot检测雷帕霉素作用后骨肉瘤细胞Fas蛋白表达的变化。结果MTT检测显示雷帕霉素可抑制人骨肉瘤细胞增殖，在雷帕霉素浓度达到20nmol／L时，抑制作用显著提高（P〈0．05），增殖抑制率分别达到31．2％和28．7％。流式细胞检测显示在雷帕霉素浓度达到20nmol/L时，其诱导骨肉瘤细胞凋亡的作用显著增强（P〈0．05）。Westernblot检测显示经雷帕霉素作用48h后的骨肉瘤细胞Fas蛋白的表达明显增强。结论雷帕霉素对人骨肉瘤细胞株U-2OS和Saos-2的增殖有抑制作用并可诱导其发生凋亡，其可能是通过上调Fas蛋白的表达而发挥作用的。

Objective To observe the effects of rapamycin （RAPA） on the proliferation and apoptosis of osteosarcoma U-2OS and Saos-2 cells and the action mechanism. Methods U-2OS and Saos-2 cells were cultured in vitro. The methyl thiazolyl tetrazoliun （MTT） and flow cytometry methods were used to detect the effects of different concentrations of rapamycin on the proliferation and apoptosis of osteosarco- ma cells. The expression of Fas protein was detected by Western blotting. Results Rapamycin could inhib- it the proliferation of human osteosarcoma cells. When the concentration of rapamycin reached 20 nmol/L, the inhibition was increased significantly,P 〈 0. 05, The inhibition rate was 31.2% and 28.7% in U-2OS and Saos-2 cells respectively. The apoptosis rate was increased notably when the concentration of rapamycin reached 20 nmol/L （P 〈 0. 05）. The protein expression of Fas was significantly increased when the rapam- ycin was given 48 h later. Conclusion Rapamycin could inhibit the proliferation and induce apoptosis of human osteosarcoma U-2OS and Saos-2 cells. It might play its role through increasing the protein expression of Fas.