Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Effect of dose-rate on nitric oxide-induced apoptosis in A375 human melanoma cells

Seo Hyun Moon¹, Min Young Kim²

¹Department of Forensic DNA, National Forensic Service, Wonju, Gangwon-do, Jeju, Republic of Korea; ²Toxicology Laboratory, Faculty of Biotechnology (Biomaterials), College of Applied Life Science, Jeju National University, Jeju, Republic of Korea.

For correspondence:- Min Kim Email: jeffmkim@jejunu.ac.kr Tel:+82647543349

Accepted: 24 March 2024 Published: 30 April 2024

Citation: Moon SH, Kim MY. Effect of dose-rate on nitric oxide-induced apoptosis in A375 human melanoma cells. Trop J Pharm Res 2024; 23(4):675-681 doi: 10.4314/tjpr.v23i4.1

© 2024 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

Abstract

Purpose: To investigate the effect of dose-rate on nitric oxide (NO)-induced apoptosis in A375 human melanoma cells.

Methods: The A375 cells were exposed to NO at a steady-state concentration of 7 μM, similar to the level estimated to occur in vivo in inflamed tissues, and delivered by diffusion through silastic tubing. Trypan blue dye exclusion assay was used to assess cell proliferation and viability. Cell cycle was studied by propidium iodide (PI) staining while Annexin V-FITC and PI assays were used to evaluate apoptosis. Western blotting assay was carried out to determine protein levels of p53, Bax, DR4, DR5, Fas (CD95), Procaspase 8, Procaspase 9 and Procaspase 3.

Results: Viability of A375 cells was reduced by 22 % after 24 h leading to extensive apoptosis and cell cycle arrest following exposure to a cumulative dose of 3360 µM/min NO. Treatment with NO stimulated p53 and triggered mitochondrial apoptotic events by inducing conformational changes in Bax. Also, activation of caspase 9 and 3, DR4, DR5, Fas (CD95) and caspase 8 appeared to be mediated concurrently by death receptor processing and downstream caspases.

Conclusion: Nitric oxide (NO) induces programmed cell death, thus indicating that it could serve as an effective inhibitor to halt the progression of melanoma or as an enhancer to improve therapeutic strategies for treatment.

Keywords: Apoptosis, A375, Cell cycle arrest, Human melanoma, Nitric oxide