Extrinsic nitric oxide donor partially reverses arginine deiminase induced cell growth inhibition through NF kappa B and Bcl-X-L

Abstract

Arginine deiminase (ADI) is known to be an inducer of apoptosis in vitro and an anti-tumor agent in vivo in some cancers. ADI causes the enzymatic depletion of arginine which may inhibit nitric oxide (NO) synthesis. However, the effect of ADI treatment on NO synthesis has not been clearly elucidated. With the goal of understanding the role of ADI in NO synthesis, we used the Ramos human lymphoma cell line, which is known to be ADI-sensitive. After determining an optimal experimental ADI concentration (0.001 U/ml), we studied the effects of ADI treatment when combined with different concentrations of the extrinsic NO donor, sodium nitroprusside (SNP) (i.e., control, ADI only, ADI with 10 mu M/ml SNP, ADI with 50 mu M/ml SNP, and ADI with 100 mu M/ml SNP). An MTT assay was used to assess cell survival after treatment, nitric oxide assays to determine nitrite levels and Western blot analysis to determine the expression of the NO mediators, NF kappa B and Bcl-X-L. Interestingly, we found that the extrinsic NO donor only partially reversed ADI-induced inhibition of cell growth in a dose-dependent pattern and resulted in an induction of NF kappa B and Bcl-X-L expression. In conclusion, we suggest that there might be an association between reversal of cell growth inhibition by extrinsic NO donor with Bcl-X-L and NF kappa B expression in ADI-treated Ramos cell.