哌立福新联合顺铂抑制骨肉瘤细胞U2-OS的活性研究

Inhibitory Effect of Perifosine Combined with Cisplatin on Human Osteosarcoma U2-OS Cells

目的探讨PI3K/Akt特异性抑制剂哌立福新协同化疗药物顺铂(DDP)对骨肉瘤细胞U2-OS增殖和凋亡的影响。方法采用四甲基偶氮唑盐(MTT)法检测哌立福新对U2-OS细胞增殖的影响;AnnexinV-FITC试剂盒检测细胞凋亡;免疫印迹法检测蛋白表达水平。结果哌立福新能够显著抑制U2-OS细胞的增殖,并下调U2-OS细胞中Akt的磷酸化水平。哌立福新明显增强顺铂对U2-OS细胞的抑制作用,并通过活化胱天蛋白酶家族成员来诱导U2-OS细胞发生凋亡。结论 PI3K/Akt抑制剂哌立福新能够显著抑制人骨肉瘤细胞U2-OS的生长;哌立福新和顺铂协同抑制U2-OS细胞的增殖并诱导其凋亡,其机制可能与共同调节胱天蛋白酶家族因子的活性相关。

Objective To detect the effect of perifosine, an inhibitor of phosphoinositol-3-kinase/Akt, on the cell growth of human osteosarcoma U2-OS ceils, and to study the molecular mechanism of the synergistic anti-proliferative effect of perifosine combined with cisplatin on U2-OS cells. Methods Cell growth inhibition was detected by 3-（4,5-dimethyhhiazol-2-yl）-2,5- diphenyl tetrazolium bromide （MTT） assay. AnnexinV-FITC apoptosis detection kit was used to detect cell apoptosis. Protein expression was examined by Western blotting. Results Akt phosphorylation was inhibited by perifosine in U2-OS cells. MTI＂ results indicated that perifosine inhibited the growth of U2-OS cells in a dose-dependent manner. Combined utilization of perifosine substantially increased cytotoxic effects of cisplatin on U2-OS ceils and induced the cell apoptosis by activating the caspase related signaling pathway. Conclusion The PI3K/Akt inhibitor perifosine can significantly inhibit the growth of U2-OS cells. Perifosine combined with cisplatin can synergistically induce apoptosis of human osteosarcoma cells. The mechanism might be related to the co- regulation of caspases activation pathways.