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观察曲古抑菌素A(TSA)对骨肉瘤细胞株143B增殖及凋亡的作用,并探讨其机制。TSA与p38抑制剂(SB203580,3μmol/L)及JNK抑制剂(SP600125,0.5μmol/L)单独或同时处理143B细胞,分别以MTT、台盼蓝染色、流式细胞术和JC-1(测定线粒体跨膜电位)法检测TSA对143B细胞的增殖、存活、周期以及凋亡的影响。应用RT-PCR、Western blot检测Bax、Bcl-2、p38/JNK表达。结果显示,TSA能够以时间和剂量依赖方式抑制143B细胞增殖,使细胞周期阻滞于G0/G1与G2/M期,并能诱导143B细胞凋亡,引起线粒体膜电位降低,促凋亡蛋白Bax表达上调,抑凋亡蛋白Bcl-2表达下调,同时使p38/JNK活化增加。p38/JNK抑制剂则能逆转TSA对Bax/Bcl-2的上调及抑制作用。研究结果揭示,TSA可以时间剂量依赖方式抑制143B细胞增殖,阻滞细胞周期,诱导细胞凋亡;其诱导细胞凋亡的机制可能与活化MAPK通路中p38和JNK的活性从而激发线粒体凋亡通路有关。
To observe the effect of trichostatin A (TSA) on the proliferation and apoptosis of osteosarcoma cell line 143B and to explore its mechanism. 143B cells were treated with TSA alone or simultaneously with p38 inhibitor (SB203580, 3μmol / L) and JNK inhibitor (SP600125, 0.5μmol / L). MTT assay, trypan blue staining, flow cytometry and JC- Mitochondrial transmembrane potential) assay TSA 143B cell proliferation, survival, cycle and apoptosis. The expression of Bax, Bcl-2 and p38 / JNK were detected by RT-PCR and Western blot. The results showed that TSA could inhibit the proliferation of 143B cells in a time and dose-dependent manner, and blocked the cell cycle in G0 / G1 and G2 / M phases, and induced the apoptosis of 143B cells, resulting in the decrease of mitochondrial membrane potential and the expression of pro-apoptotic protein Bax The expression of Bcl-2 was down-regulated while the activation of p38 / JNK was increased. p38 / JNK inhibitor could reverse the up-regulation and inhibition of Bax / Bcl-2 by TSA. The results showed that TSA can inhibit the proliferation of 143B cells in a time-and dose-dependent manner, arresting the cell cycle and inducing apoptosis. The mechanism of TSA induction may be related to the activation of p38 and JNK in the MAPK pathway and the activation of mitochondrial apoptotic pathway .