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
Journal Title
Journal ISSN
Volume Title
Publisher
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.
Description
Keywords
Prostate Carcinogenesis , Heterocyclic Amine 2-Amino-1-Methyl- 6-Phenylimidazo[4,5-B]Pyridine