曲古抑菌素A(TSA)对骨肉瘤细胞株143B增殖和凋亡的作用及其机制  

Effects and Potential Mechanisms of Trichostatin A(TSA) on the Proliferation and Apoptosis of Osteosarcoma Cell Line 143B

作　　者：杨阳[1] 严树涓[1] 夏菁[2] 石庆强 姚娟[1] 幸艺芳 翁亚光[1] 左国伟[1] 

机构地区：[1]重庆医科大学检验医学院,临床检验诊断学教育部重点实验室,重庆市重点实验室,重庆400016 [2]重庆医科大学基础医学院,组织学与胚胎学教研室,重庆400016

出　　处：《中国细胞生物学学报》2014年第2期169-176,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81102035);重庆市教委基金(批准号:KJ110328)资助的课题~~

摘　　要：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。This paper studied the effects of Trichostatin A （TSA） on the proliferation and apoptosis of osteosarcoma cell line143B and its potential mechanisms. 143B cells were treated with indicated doses of TSA in the presence or absence of p38 inhibitor （SB203580, 3 pmol/L） and JNK inhibitor （SP600125, 0.5 μmol/L）, and then the proliferation activity of cell line was determined with MTT assay and trypan blue staining. Cell apoptotic and cell cycle were observed by flow cytometry. The mitochondrial membrane potential detection was observed with JC-1 assay. The expression levels of Bax, Bcl-2, p38/JNK were determined by RT-PCR and Western blot. The results showed that TSA could significantly suppress the proliferation of 143B in a time- and dose-dependent manner with MTT/trypan blue staining analysis. The apoptotic rate was increased with TSA treatment, and the cell cycle arrest was observed （staying in G0/Ga and G/M）, and the decrease of mitochondrial membrane potential detection was also visible. RT-PCR and Western blot showed that the expressions of Bax, Bcl-2 and p38/JNK were increased. p38/JNK inhibitors could reverse the effects of TSA on the expression level of BaxfBcl-2. These data demonstrated that TSA could inhibit the proliferation of 143B cell in a time- and dose-dependent manner. TSA cloud also induce apoptosis and cell cycle arrest of 143B with different concentrations. The mechanisms of TSA induced apoptosis may attribute to the stimulation of mitochondrial apoptosis pathway by activating p38/MAPK and JNK/MAPK pathways.

关 键 词：曲古抑菌素A 骨肉瘤 增殖 凋亡 

分 类 号：R738.1[医药卫生—肿瘤]