Microarray study of mechanism of trichostatin a inducing apoptosis of Molt-4 cells
Zhenya Hong1, Zhiqiang Han, Min Xiao
1Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China. hongzhenya@eyou.com
Abstract:
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.
Insights
Trichostatin A (TSA) induces apoptosis in leukemia cells by altering gene expression, but spares normal cells. This histone deacetylase inhibitor shows promise for cancer therapy by selectively targeting cancer cell growth and survival mechanisms.
Area of Science:
- Molecular Biology
- Cancer Research
- Cell Biology
Background:
- Histone deacetylase (HDAC) overexpression is linked to various cancers.
- HDAC inhibitors like trichostatin A (TSA) induce cancer cell apoptosis, but mechanisms are unclear.
Purpose of the Study:
- To investigate the global gene expression profiles of human leukemia Molt-4 cells treated with TSA.
- To elucidate the mechanism of TSA-induced apoptosis and growth inhibition in leukemia cells.
Main Methods:
- Flow cytometry, MTT assay, and DNA laddering to assess apoptosis.
- Microarray analysis, RT-PCR, and Western blotting to detect gene and protein expression changes.
Main Results:
- TSA induced dose- and time-dependent apoptosis in Molt-4 cells, sparing normal peripheral blood mononuclear cells (PBMCs).
- Microarray analysis revealed significant upregulation and deregulation of genes involved in cell growth, differentiation, and survival.
- Key genes like STAT5A and MYC were significantly downregulated post-TSA treatment.
Conclusions:
- TSA exhibits selective growth-inhibiting and apoptosis-inducing effects on leukemia cells.
- The mechanism involves modulation of pro-proliferation and anti-apoptosis gene expression profiles.
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