TRAIL联合顺铂对横纹肌肉瘤细胞Fas表达和caspase3活性的影响

Effect of Fas and caspase3 activation in human rhabdomyosarcoma cells by combining TRAIL with cisplatin

目的:探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)和顺铂(DDP)协同抑制横纹肌肉瘤(rhabdomyosarcoma,RMS)细胞生长和诱导凋亡的作用。方法:将不同浓度的TRAIL和DDP作用于人RD细胞,通过噻嗟蓝(MTT)比色法、丫啶橙染色及流式细胞仪分析细胞的增殖、形态学、细胞凋亡、Fas蛋白表达和caspase3活性改变。结果:浓度分别为1.0、10.0和100.0μg/L的TRAIL作用于RD细胞后,细胞生长抑制率(CI)分别为18.9%、20.8%和43.5%;浓度分别为1.0、5.0和10.0mg/L的DDP作用于RD细胞后,CI分别为9.8%、23.4%和43.8%。而浓度为100μg/L的TRAIL与浓度为5mg/L的DDP联合作用时,CI明显增加(66.4%),Fas蛋白的表达及caspase3的活性也明显提高。荧光显微镜下可见明显的凋亡细胞形态特征。结论:TRAIL和DDP联合应用对诱导RMS细胞凋亡具有明显的协同效果,这一作用与增加Fas和caspase3活性有关。

OBJECTIVE： To study the synergistic induction of apoptosis by the combination of TRAIL and cisplatin in rhabdomyosarcoma ceils. METHODS： Rhabdomyosarcoma cells were treated with TRAIL, cisplatin for 3 days, respectively. The eytotoxieity was observed by MTT assay. The apoptotic rates and changes of Fas and caspase3 activation were shown by flow cytometry （FCM）. The obvious morphological changes in rhabdomyosarcoma cells were comfirmed by a fluorescence microscope. RESULTS： After treated with TRAIL（1. 0, 10.0, 100.0 μg/L）, the eytotoxicity indexes of rhabdomyosarcome cells were 18.9%, 20.8%, 43.5%, respectively. with cisplatin（1.0, 5.0, 10.0 mg/L）, the cytotoxicity indexes were 9.8%, 23.4%, 43.8%, respectively. TRAIL and cisplatin treatment used simultaneously, the cytotoxicity index increased obviously, Fas and caspase3 activation increased significantly, which were paralleled by the apoptotic rates. The ob vious apotosis morphological changes in rhabdomyosarcoma cells were shown by a fluorescence microscope. CONCLUSION： TRAIL and cisplatin are able to kill rhabdomyosarcoma cells, respctively, while TRAIL with cisplatin together applied can have synergistic effect on rhabdomyosarcoma cells by increasing the caspase3 activity and suppressing mitochondrial membrane potential.