alexa Effect of 2-Methoxyestradiol on Dephosphorylation of Ne
ISSN: 2161-0495

Journal of Clinical Toxicology
Open Access

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Research Article

Effect of 2-Methoxyestradiol on Dephosphorylation of Neuronal Nitric Oxide Synthase in Osteosarcoma 143B Cells. An in vitro study

Gorska M1*, Kuban-Jankowska A1, Antoniewicz J2 and Wozniak M1

1Department of Medical Chemistry, Medical University of Gdansk, Gdansk 80-211, Poland

2Department of Histology, Medical University of Gdansk, Gdansk, Poland

*Corresponding Author:
Gorska M
Department of Medical Chemistry
Medical University of Gdansk
Gdansk 80-211, Poland
Tel: +48-349-14-50
Fax: +48-349-14-56
E-mail: [email protected]

Received date: February 28, 2015; Accepted date: March 26, 2015; Published date: March 30, 2015

Citation: Gorska M, Kuban-Jankowska A, Antoniewicz J, Wozniak M (2015) Effect of 2-Methoxyestradiol on Dephosphorylation of Neuronal Nitric Oxide Synthase in Osteosarcoma 143B Cells. An in vitro study. J Clin Toxicol 5:240. doi: 10.4172/2161-0495.1000240

Copyright: © 2015 Gorska M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 

Abstract

Objective: 2-methoxyestradiol, an endogenous metabolite of anticancer agent, has been evaluated in ongoing advanced phases of clinical trials as an anticancer agent. The aim of the study was to determine the impact of 2- methoxyestradiol on phosphorylation status of neuronal nitric oxide synthase at position of serine 847. We determined also influence of the compound on the gene and protein expressions, phosphorylation status of PP2A phosphatase.

Methods: In the current study we used western blotting, real time PCR, and ELISA assays.

Results: We demonstrated that 2-methoxyestradiol decreases the level of neuronal nitric oxide synthase phosphorylated at position of serine 847 referred to as inactive pool of enzyme. The expression of PP2CA gene was down-regulated by 2-methoxyestradiol at concentration of 1 μM. 10 μM did not statistically significant impact on phosphatase gene expression. Moreover, 1 μM 2-methoxyestradiol up-regulated protein level of PP2A while 10 μM did not exert any effect.

Conclusions: 2-methoxyestradiol decreased an inactive pool of neuronal nitric oxide synthase. Additionally, in a concentration-dependent manner it impacts on the gene and protein expressions, as well as phosphorylation status of PP2A-α phosphatase. Increasing activity of neuronal nitric oxide synthase by reversible phosphorylation may constitute an important tool resulting in osteosarcoma cell death.

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