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Research article

ATR-FTIR spectroscopy detects alterations induced by organotin(IV) carboxylates in MCF-7 cells at sub-cytotoxic/-genotoxic concentrations

Muhammad S Ahmad^1,2 , Bushra Mirza² , Mukhtiar Hussain³ , Muhammad Hanif³ , Saqib Ali³ , Michael J Walsh¹ and Francis L Martin¹

¹  Lancaster Environment Centre, Lancaster University, Bailrigg, Lancaster LA1 4YQ, UK

²  Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan

³  Department of Chemistry, Quaid-i-Azam University, Islamabad, Pakistan

author email corresponding author email

PMC Biophysics 2008, 1:3doi:10.1186/1757-5036-1-3

Published:	5  November  2008

Abstract

The environmental impact of metal complexes such as organotin(IV) compounds is of increasing concern. Genotoxic effects of organotin(IV) compounds (0.01 μg/ml, 0.1 μg/ml or 1.0 μg/ml) were measured using the alkaline single-cell gel electrophoresis (comet) assay to measure DNA single-strand breaks (SSBs) and the cytokinesis-block micronucleus (CBMN) assay to determine micronucleus formation. Biochemical-cell signatures were also ascertained using attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy. In the comet assay, organotin(IV) carboxylates induced significantly-elevated levels of DNA SSBs. Elevated micronucleus-forming activities were also observed. Following interrogation using ATR-FTIR spectroscopy, infrared spectra in the biomolecular range (900 cm^-1 – 1800 cm^-1) derived from organotin-treated MCF-7 cells exhibited clear alterations in their biochemical-cell fingerprint compared to control-cell populations following exposures as low as 0.0001 μg/ml. Mono-, di- or tri-organotin(IV) carboxylates (0.1 μg/ml, 1.0 μg/ml or 10.0 μg/ml) were markedly cytotoxic as determined by the clonogenic assay following treatment of MCF-7 cells with ≥ 1.0 μg/ml. Our results demonstrate that ATR-FTIR spectroscopy can be applied to detect molecular alterations induced by organotin(IV) compounds at sub-cytotoxic and sub-genotoxic concentrations. This biophysical approach points to a novel means of assessing risk associated with environmental contaminants.

PACS codes: 87.15.-v, 87.17.-d, 87.18.-h