伏立诺他联合顺铂对人骨肉瘤细胞增殖与凋亡的作用

Effects of vorinostat in combination with cisplatin on proliferation and apoptosis of human osteosarcoma cells

目的:探讨组蛋白去乙酰化酶抑制剂(histone deacetylase inhibitor,HDACi)伏立诺他联合顺铂对骨肉瘤U2OS细胞增殖、细胞周期和细胞凋亡的影响及其可能的作用机制。方法:采用MTS法检测伏立诺他联合顺铂对U2OS细胞增殖的影响,FCM法检测伏立诺他联合顺铂对U2OS细胞周期分布、线粒体膜电位(mitochondrial membrane potential,MMP)势能和细胞凋亡的影响,蛋白质印迹法检测伏立诺他联合顺铂对U2OS细胞中caspase-3、caspase-8、caspase-9、多聚ADP-核糖聚合酶(poly ADP-ribose polymerase,PARP)、c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)/磷酸化JNK(phospho-JNK,p-JNK)蛋白表达的影响。结果:伏立诺他联合顺铂可显著抑制U2OS细胞的增殖(P<0.05),并将细胞阻滞于S期(P<0.05)。同时,伏立诺他联合顺铂可降低U2OS细胞的MMP势能(P<0.01),伏立诺他联合顺铂处理组U2OS细胞中caspase-8、caspase-9、PARP和p-JNK蛋白表达水平上调(P值均<0.05)。结论:伏立诺他联合顺铂通过JNK/caspase信号通路诱导骨肉瘤细胞周期阻滞及凋亡,具有显著抑制骨肉瘤细胞增殖的作用。

Objective： To investigate the effects of histone deacetylase inhibitor （HDACi） vorinostat in combination with cisplatin on the proliferation, cell cycle and apoptosis of human osteosarcoma U2OS cells, and to explore the possible mechanisms.Methods： After the treatment with vorinostat and cisplatin, the proliferation of U2OS cells was detected by MTS method, while the cell cycle distribution, mitochondrial membrane potential and apoptosis of U2OS cells were detected by FCM. Furthermore,the effects of vorinostat in combination with cisplatin on the expressions of caspase-3, caspase-8, caspase-9, poly ADP-ribose polymerase （PARP）, c-Jun N-terminal kinase UNK） and phospho-JNK （p-JNK） proteins in U2OS cells were detected by Western blotting,Results： The proliferation of U2OS cells was inhibited by vorinostat in combination with cisplatin （P 〈 0.05）, and the cell cycle was arrested at S phase （P 〈 0.05）. The mitochondrial membrane potential of U2OS cells was decreased （P 〈 0.01）, and the apoptosis of U2OS cells was induced by vorinostat in combination with cisplatin （P 〈 0.05）. The expression levels of caspase-3, caspase-8, caspase-9, PARP and p-JNK proteins in U2OS cells after the treatment with vorinostat and cisplatin were down-regulated （all P 〈 0.05）.Conclusion： Vorinostat in combination with cisplatin can induce the cell cycle arrest and apoptosis of osteosarcoma cells as well as suppress the proliferation. The mechanism may be associated with JNK/caspase signaling pathway.