Hong Zhenya, Han Zhiqiang, Xiao Min, Yang Yang, Xia Xi, Zhou Jianfeng
Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2009 Aug;29(4):445-50. doi: 10.1007/s11596-009-0411-y. Epub 2009 Aug 7.
Histone deacetylase was overexpressed in a variety of cancers and was closely correlated with oncogenic factors. The histone deacetylase inhibitor, trichostatin A (TSA) was shown to induce apoptosis in many cancer cells. However, the mechanism of TSA on induction of cancer cells apoptosis is poorly understood. This study was designed to characterize the global gene expression profiles before and after treatment of human leukemia cell line Molt-4 with TSA. Flow cytometry, MTT and DNA ladder were used to observe the effect of TSA on the apoptosis of MOLT-4 cells and normal human peripheral blood mononuclear cells (PBMC). Microarray, reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to detect the difference of gene and protein expressions of Molt-4 cells after incubation of the cells with TSA. The results showed that TSA could induce Molt-4 apoptosis in dose- and time-dependent manners but spared PBMCs. Microarray analysis showed that after incubation with TSA for 9 h, 310 genes were upregulated and 313 genes were deregulated. These genes regulate the growth, differentiation and survival of cells. Among these genes, STAT5A was down-regulated by 80.4% and MYC was down-regulated by 77.3%. It was concluded that TSA has definite growth-inhibiting and apoptosis-inducing effects on Molt-4 cells in time- and dose-dependent manners, with weak cytotoxic effects on PBMCs at the same time. The mechanism of TSA selectively inducing apoptosis and inhibiting growth may be ascribed to the changes of pro-proliferation genes and anti-apoptosis genes.
组蛋白去乙酰化酶在多种癌症中过表达,且与致癌因素密切相关。组蛋白去乙酰化酶抑制剂曲古抑菌素A(TSA)已被证明可诱导多种癌细胞凋亡。然而,TSA诱导癌细胞凋亡的机制尚不清楚。本研究旨在表征TSA处理人白血病细胞系Molt-4前后的全基因组表达谱。采用流式细胞术、MTT法和DNA梯状条带分析观察TSA对MOLT-4细胞和正常人外周血单个核细胞(PBMC)凋亡的影响。利用基因芯片、逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹法检测TSA作用于Molt-4细胞后基因和蛋白表达的差异。结果显示,TSA能以剂量和时间依赖性方式诱导Molt-4细胞凋亡,但对PBMC无影响。基因芯片分析显示,TSA作用9小时后,有310个基因上调,313个基因下调。这些基因调控细胞的生长、分化和存活。其中,信号转导子和转录激活子5A(STAT5A)下调80.4%,原癌基因MYC下调77.3%。研究得出结论,TSA对Molt-4细胞具有明确的时间和剂量依赖性生长抑制及凋亡诱导作用,同时对PBMC的细胞毒性较弱。TSA选择性诱导凋亡和抑制生长的机制可能归因于促增殖基因和抗凋亡基因的变化。