Proteomic Analysis Of Prostate Carcinogenesis Induced By Heterocyclic Amine 2-Amino-1-Methyl- 6-Phenylimidazo[4,5-B]Pyridine (Phip) Using In Vitro Model

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Date
2019-01
Authors
Saifuddin, Siti Nazmin
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Universiti Sains Malaysia
Abstract
2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is the most abundant type of carcinogenic heterocyclic amine produced in cooked meat. This study aims to investigate the carcinogenic effect of human physiologically relevant concentration of PhIP on human prostate epithelial cell line, RWPE-1, at cellular and protein levels in prostate carcinogenesis. RWPE-1 cells were exposed to 10-7, 10-8, 10-9 or 10-10 M PhIP, diluted in 0.003%, 0.025% or 0.25% DMSO, with or without metabolic activation system. The methodologies used in this study were MTS assay for cytotoxicity and cell proliferation tests, Alamar Blue assay for cell proliferation test, cell transformation assay, invasion assay and shotgun proteomic. In cytotoxicity and cell proliferation assays, results showed that the viability and proliferation of RWPE-1 cells treated with PhIP at all doses in all DMSO concentrations in the presence of metabolic activation system were increased depending on the vehicle concentration, where the lower the concentration of the vehicle used, the higher the cellular responses observed. In the cell transformation assay conducted using the same experimental settings as the previous assays, the results showed that all PhIPtreated groups with metabolic activation system exhibited neoplastic transformation characteristic as these transformed cells showed anchorage-independent growth growing above the agar layer and formed colonies in soft agar.
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Keywords
Prostate Carcinogenesis , Heterocyclic Amine 2-Amino-1-Methyl- 6-Phenylimidazo[4,5-B]Pyridine
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