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Updated: Jul 18, 2026
Assays for Validating Histone Acetyltransferase Inhibitors
Published on: August 6, 2020
[Anti-proliferative effect of histone deacetylase inhibitors on murine embryonic stem cells]
Abstract:
The effect of histone deacetylase (HDAC) inhibitors trichostatin A (TSA) and sodium butirate (NaBut) on the proliferation of murine embryonic stem cells (MESC) was studied. Both agents suppressed the population growth and clonability of MESC. Flow cytometry analysis showed a decrease in the amount of S-phase cells upon treatment with HDAC inhibitors. TSA treatment caused a decrease in mRNA level of such positive cell cycle regulators as cyclins D1, A, c-myc, cdc25A, and induced transcription of negative regulators of the cell cycle--p21(Wafl) and p57(kip2). Also, HDAC inhibitors decreased the level of e2f-dependent transcription, with the concominant reduction of mRNA level of e2fl gene. HDAC inhibitors also affected the survival of MESC. A 2 day TSA treatment resulted in massive detachment and cell death, as confirmed by DNA laddering and MTT assay. Treatment with TSA for 2 and 5 days did not induce SAPbetaGAL, activity and p16(ink4a) transcription, i.e., characteristic features of senescent fibroblasts. In summary, HDAC inhibitors decrease the rate of proliferation affecting cell cycle and viability of MESC. We conclude that MESC are unable to realize a sustainable block of the cell cycle upon treatment with HDAC inhibitors.
Insights
Histone deacetylase (HDAC) inhibitors like TSA and NaBut reduce the proliferation and viability of murine embryonic stem cells (MESC). These HDAC inhibitors disrupt the cell cycle, leading to decreased S-phase cells and increased cell death.
Area of Science:
- Stem cell biology
- Epigenetics
- Cell cycle regulation
Context:
- Murine embryonic stem cells (MESCs) are crucial for developmental biology research.
- Histone deacetylase (HDAC) inhibitors are emerging as therapeutic agents with diverse cellular effects.
- Understanding the impact of HDAC inhibitors on stem cell function is vital for regenerative medicine and cancer research.
Purpose:
- To investigate the effects of HDAC inhibitors, specifically trichostatin A (TSA) and sodium butyrate (NaBut), on the proliferation, cell cycle progression, and survival of MESCs.
Summary:
- Treatment with TSA and NaBut suppressed MESC proliferation and clonability.
- HDAC inhibitors reduced the proportion of cells in S-phase and altered the expression of key cell cycle regulators (cyclins, c-myc, p21, p57).
- TSA induced significant MESC death, evidenced by DNA laddering and MTT assay, without triggering senescence markers like p16ink4a.
Impact:
- HDAC inhibitors impair MESC proliferation and viability by disrupting cell cycle control.
- MESCs exhibit an inability to establish a sustained cell cycle arrest in response to HDAC inhibition.
- These findings highlight the critical role of HDACs in maintaining stem cell homeostasis and suggest potential therapeutic vulnerabilities.
