Effects of Methionine Gamma Lyase-Deaminase on Human Colorectal Carcinoma Cells

Nova Southeastern University, Davie, Florida, USA

Objective. The objective of this study is to demonstrate the effects of methionine gamma lyase-deaminase (Mgld) on colorectal carcinoma cell survival using MTT assay. Background. Mgld is in enzyme found in bacteria and protozoa including, Porphyromonas gingivalis. Mgld has the ability to catabolize methionine, an enzyme function that is absent in mammals. This enzyme function is of interest in methionine/S-adenosyl methionine (SAM) dependent cancer cells given the dysregulation of DNA methylation patterns that occur compared to normal mammalian cells. Methods. In this study, Porphyromonas gingivalis Mgld cloned into a cytoplasmic and nuclear plasmid vector was transfected into the human colorectal carcinoma T84 cell line using Lipofectamine 3000. The effects of cytoplasmic Mgld and nuclear Mgld were assessed in comparison to control non-transfected cells. Cell survival was assessed with an MTT assay at 570 nm as an indicator of metabolic functions of the live cells/mitochondria. Results. Results indicate that nuclear Mgld transfection causes most significant inhibition of metabolic activity in T84 colorectal carcinoma cells with a 19% decrease in absorbance compared to the control. Conclusion. Due to its effects on cellular survival, further studies should be conducted to evaluate and investigate the metabolic implications of nuclear Mgld on T84 colorectal carcinoma cells.