Tropical Journal of Pharmaceutical Research
Pharmacotherapy Group, Faculty of Pharmacy, University of Benin, Benin City, Nigeria
Vol. 15, No. 8, 2016, pp. 1595-1603
Bioline Code: pr16209
Full paper language: English
Document type: Research Article
Document available free of charge
Tropical Journal of Pharmaceutical Research, Vol. 15, No. 8, 2016, pp. 1595-1603
© Copyright 2016 - Tropical Journal of Pharmaceutical Research
Cellular inactivation of nitric oxide induces p53-dependent apoptosis in human melanoma cells|
Moon, Seo Hyun; Cho, Myung Haing & Kim, Min Young
Purpose: To examine the role of endogenous nitric oxide (NO•) and influence of p53 status during
apoptosis induced by a selective iNOS inhibitor, N-[(3-aminomethyl) benzyl] acetamidine (1400W),
and/or an NO• scavenger carboxy-PTIO (c-PTIO) in two isogenic human melanoma cell lines, wild-type
p53 (A375) and p53 mutant (SK mel-28) cells.
Methods: 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) and Annexin V/propidium
iodide assay were used to test for antiproliferation and apoptosis, respectively. Griess and reverse
transcription-polymerase chain reaction (RT-PCR) reactions were carried out to assay NO• production
and the mRNA levels of inhibitors of apoptosis (IAP).
Results: c-PTIO and 1400W, alone or in combination, inhibited cell growth and promoted apoptosis via
sub-G1 cell cycle arrest mediated by decrease in NO•. Apoptosis was delayed and greatly reduced in
magnitude in SK mel-28 cells, underscoring the importance of p53 modulation of the response. In both
cell types, apoptosis induced by iNOS inhibition and/or NO• depletion was blocked by an exogenous
NO• donor, sodium nitroprusside. It was also found that inhibitors of apoptosis family (survivin, XIAP and
cIAP1) were significantly depressed, which appear to play an important role in the regulation of p53-
mediated apoptotic response under these conditions.
Conclusion: The data obtained provide insight into the mechanism of cell proliferation action of
endogenous NO•, based on p53 status, and indicate manipulation of iNOS may offer exciting
opportunities to improve the effectiveness of melanoma treatment.
Apoptosis; Human melanoma cells; Inducible nitric oxide synthase; p53
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