TRAIL与顺铂协同诱导横纹肌肉瘤细胞凋亡时Fas和cFLIP表达的改变及其意义

Changes and significance of Fas and cFLIP in apoptosis of human rhabdomyosarcoma cells induced by combination of TRAIL and cisplatin

目的 :探讨肿瘤坏死因子相关凋亡诱导配体 (TRAIL)及其基因Ad/GT -TRAIL和顺铂 (DDP)联合应用对人横纹肌肉瘤细胞生长抑制和诱导凋亡作用 ,并分析细胞表面Fas蛋白和细胞内cFLIPmRNA表达对凋亡影响。方法 :将TRAIL(10 0 μg/L)、Ad/GT -TRAIL和顺铂作用于培养的人RD横纹肌肉瘤细胞 ,通过MTT比色法、流式细胞仪检测细胞凋亡和Fas蛋白表达、RT -PCR检测cFLIPmRNA表达 ,观察和分析TRAIL及其基因Ad/GT -TRAIL对横纹肌肉瘤细胞的作用及和顺铂协同作用的机制。结果 :Ad/GT -TRAIL和 10 0 μg/LTRAIL对横纹肌肉瘤细胞的生长抑制率分别为 5 2 5 %和 4 3 5 % ,凋亡诱导率为 12 95 %和 10 2 6 % ,联合应用顺铂 ,生长抑制率和凋亡诱导率均显著高于单独应用 (P <0 0 5 ) ,FCM分析显示联合应用提高了Fas蛋白的表达 ,RT -PCR显示cFLIPmRNA表达下降 ,与联合应用组细胞凋亡率增加相一致。结论 :TRAIL和Ad/GT -TRAIL能有效诱导横纹肌肉瘤细胞的凋亡从而抑制横纹肌肉瘤细胞的生长 。

AIM: To investigate the synergistic induction of apoptosis in rhabdomyosarcoma cells by the combination of TRAIL or TRAIL gene with cisplatin. METHODS: Rhabdomyosarcoma cells were treated with TRAIL, (Ad/GT)-TRAIL, cisplatin, respectively or the combination for 3 days. The cytotoxicity was observed by MTT assay. The apoptotic rates and the expression rates of Fas protein were measured by flow cytometry (FCM). The expression of cFLIP mRNA was determined by RT-PCR. RESULTS: Rhabdomyosarcoma cells were treated with Ad/ GT-TRAIL and TRAIL (100.0 μg/L), the cytotoxicity index were 52.5% and 43.5%, the percentage of apoptotic cells were 12.95% and 10.26%, respectively. Combined with cisplatin, the cytotoxicity index and the percentage of apoptotic cells were increased significantly (P<0.05). The expression of Fas protein in rhabdomyosarcoma cells was up-regulated and the expression of cFLIP was down-regulated with cisplatin, which were paralleled by the apoptotic rates. CONCLUSION: Combinatiion of Ad/GT-TRAIL or TRAIL and cisplatin has synergistic apoptosis-inducing effects on rhabdomyosacoma cells. [