Valproic acid (Synonyms: NSC 93819, 2-Propylvaleric Acid, Valproate, VPA)

In vitro Kinase assays

For in vitro HDAC assays, myc epitope-tagged HDAC1 was transfected into HeLa cells and immunoprecipitated. Immunoprecipitates were washed, resuspended in HD buffer (20 mM Tris-HCl, pH 8.0, 150 mM NaCl, 10% glycerol), and stored as frozen aliquots. HDAC1 was then added to a tube containing 40,000 cpm 3H-labeled acetylated histones (purified from HeLa cells) in 200 μl of HD buffer ±valproic acid,. After rotation for 2 h at 37 °C, the reaction was stopped by the addition of 50 μl of stop solution and released 3H-labeled acetic acid was extracted and analyzed by scintillation counting.

Cell lines

Neuro2A cells, human ovarian cancer cell line SKOV3

Preparation method

Soluble in DMSO. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reacting condition

0.5–5 mM for 24 h; or 4 mM for 48 h

Applications

Valproic acid induced hyperacetylation of endogenous histones and inhibited nuclear HDAC activity in Neuro2A cells. Moreover, valproic acid inhibited cell proliferation, and induced apoptosis of SKOV3 cells in a dose- and time- dependent manner.

Animal models

human ovarian cancer model transplanted subcutaneously in nude mice

Dosage form

500mg/kg/day, intraperitoneal injection, for 30 days

Applications

Valproic acid induced growth inhibition of human ovarian cancer transplanted subcutaneously in nude mice. Moreover, valproic acid (50 mg/kg, IV infusion) decreased pro-inflammatory cytokine gene expression in a canine endotoxemia model in vivo.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

1. Phiel, C. J., Zhang, F., Huang, E. Y., Guenther, M. G., Lazar, M. A. and Klein, P. S. (2001) Histone deacetylase is a direct target of valproic acid, a potent anticonvulsant, mood stabilizer, and teratogen. J Biol Chem. 276, 36734-36741

2. Shan, Z., Feng-Nian, R., Jie, G. and Ting, Z. (2012) Effects of valproic acid on proliferation, apoptosis, angiogenesis and metastasis of ovarian cancer in vitro and in vivo. Asian Pac J Cancer Prev. 13, 3977-3982

3. Song, R., Yu, D., Yoon, J. and Park, J. (2015) Valproic acid attenuates the expression of pro-inflammatory cytokines lipopolysaccharide-treated canine peripheral blood mononuclear cells (in vitro) and in a canine endotoxemia model (in vivo). Vet Immunol Immunopathol. 166, 132-137