Biochemical and Biophysical Research Communications
Regulation of apoptosis by resveratrol through JAK/STAT and mitochondria mediated pathway in human epidermoid carcinoma A431 cells
Section snippets
Materials and methods
Chemicals. Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum, antibiotic–antimycotic mixture (10,000 U/ml penicillin G, 10,000 μg/ml streptomycin sulfate, and 25 μg/ml amphoterecin B), trypsin and other chemicals of culture grade were procured from Gibco Life Sciences (India). Resveratrol (>99% pure), agarose, MTT (3-[4,5-dimethyl-2-yl]-2,5-diphenyltetrazolium bromide), dichlorodihydrofluorescein diacetate dye (DCFH-DA), rhodamine 123, propidium iodide (PI), Protease inhibitor Cocktail
Results and discussion
Preliminary screening was performed to assess the effect of resveratrol on cellular proliferation and cell viability at different time intervals using the MTT assay. There was dose- (10–150 μM) and time-dependent (24, 48, and 72 h) reductions in the viability of A431 cells treated with resveratrol (Fig. 1A). The IC50 value for growth inhibition was obtained at 40 μM treatment after 24 h incubation. Based on this observation, we selected doses (40 and 60 μM) for further mechanistic studies over a
Acknowledgments
The authors thank Dr. Ashwani Kumar (Director, Indian Institute of Toxicology Research, Lucknow) for his keen interest in the study. The authors also thank the Indian Council of Medical Research, New Delhi, for providing fellowships to Sahdeo Prasad and Jasmine George.
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