Trichostatin A

Biological Activity

Trichostatin A (TSA) is an antifungal antibiotic derived from Streptomyces that inhibits mammalian histone deacetylase (HDAC). TSA inhibits the eukaryotic cell cycle during the beginning of the growth stage. Trichostatin A (TSA) can be used to alter gene expression by interfering with the removal of acetyl groups from histones (histone deacetylases, HDAC) and therefore altering the ability of DNA transcription factors to access the DNA molecules inside chromatin. It is a member of a larger class of histone deacetylase inhibitors (HDIs or HDACIs) that have a broad spectrum of epigenetic activities. Trichostatin A has also been shown to inhibit both G1- and G2-phases of the mammalian cell cycle and has been tested for use as a potential anticancer agent.

Protocol

Cell Experiment
Cell lines	MCF-7, T-47D, ZR-75-1, BT-474, MDA-MB-231, MDA-MB-453, CAL 51, and SK-BR-3 cell lines
Preparation method	Cell Proliferation Assay. Stock cultures of breast cancer cell lines MCF-7, T-47D, ZR-75-1, BT-474, MDA-MB-231, MDA-MB-453, CAL 51, and SK-BR-3 (American Type Culture Collection, Rockville, MD) were grown in DMEM containing 10% (v/v) FCS, 2 mM L-glutamine, 100 units/ml penicillin, and 100 mg/ml streptomycin at 37°C in 5% CO2 humidified atmosphere. Cells were counted in a hemocytometer after detachment using 0.25% (w/v) trypsin in Dulbecco’s PBS without Ca21 or Mg21 (DPBS; Sigma-Aldrich) containing 0.02% (w/v) EDTA. Viability was determined by trypan blue exclusion. For each cell line, cells were seeded in 96-well microtiter plates at optimal densities determined in prior experiments to ensure exponential growth for the duration of the assay. After a 24-h preincubation, growth medium was replaced with experimental medium containing TSA at final concentrations ranging from 10-12 M to 10-5 M in log dilutions and 0.1% (v/v) ethanol, or growth medium containing 0.1% (v/v) ethanol as a vehicle control. After 96 h incubation, cell proliferation was estimated using the sulforhodamine B colorimetric assay (11), and the results are expressed as the mean 6 SD for six replicates as a percentage of vehicle control (taken as 100%).
Concentrations	10-12 M to 10-5 M
Incubation time	96 h

Animal Experiment
Animal models	Inbred virgin female (Ludwig/Wistar/ Olac) rats bearing tumors induced with NMU model
Formulation	DMSO
Dosages	500 µg/kg TSA in50 µl DMSO twice weekly for 4 weeks
Administration	s.c. injection

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Chemical Information

Customer Product Validations & Biological Datas

	Source	Oncotarget (2017). Figure 2. Trichostatin A
	Method	Western blotting
	Cell Lines	RLE-6TN cells
	Concentrations	100 nM
	Incubation Time	72 h
	Results	Snail was significantly increased in irradiated group which was effectively modulated by the treatment of TSA, as we observed from the data of western blot
	Rating

	Source	Oncotarget (2017). Figure 1. Trichostatin A
	Method	Western blotting
	Cell Lines	RLE-6TN cells
	Concentrations	100 nM
	Incubation Time	72 h
	Results	TSA led to a significant modification in the protein and gene expressions of both E-cadherin and α-SMA in cells collected at 72 h after irradation.
	Rating

References

Trichostatin A selectively suppresses the cold-induced transcription of the ZmDREB1 gene in maize.
Hu et al. PLoS One. 2011;6(7):e22132. PMID: 21811564.

Trichostatin A attenuates airway inflammation in mouse asthma model.
Choi et al. Clin Exp Allergy. 2005 Jan;35(1):89-96. PMID: 15649272.

Trichostatin A induces differential cell cycle arrests but does not induce apoptosis in primary cultures of mitogen-stimulated rat hepatocytes.
Papeleu et al. J Hepatol. 2003 Sep;39(3):374-82. PMID: 12927923.

Trichostatin A is a histone deacetylase inhibitor with potent antitumor activity against breast cancer in vivo.
Vigushin et al. Clin Cancer Res. 2001 Apr;7(4):971-6. PMID: 11309348.