title: Distinguishing nuclei-specific benzo[a]pyrene-induced effects from whole-cell alterations in MCF-7 cells using Fourier-transform infrared spectroscopy.
abstract: Exposure to chemicals such as benzo[a]pyrene (B[a]P) can generate intracellular toxic mechanisms. Fourier-transform infrared (FTIR) spectroscopy is a novel approach that allows the non-destructive analysis of underlying chemical bond alterations in patho-physiological processes. This study set out to examine whether B[a]P-induced whole cell alterations could be distinguished from effects on nuclei of exposed cells. Using attenuated total reflection FTIR (ATR-FTIR) spectroscopy, alterations in nuclei isolated from B[a]P-treated MCF-7 cells concentrated either in G0/G1- or S-phase were observed. B[a]P-induced effects in whole-cells included alterations to lipids, DNA and protein spectral regions. Absorbance areas for protein and DNA/RNA regions in B[a]P-treated whole cells differed significantly (P<0.0001) from vehicle controls and these observations correlated with alterations noted in isolated nuclei. Our findings provide evidence that FTIR spectroscopy has the ability to identify specific chemical-induced alterations.
authors.affiliation: Centre for Biophotonics, Lancaster Environment Centre, Lancaster University, Bailrigg, Lancaster LA1 4YQ, UK.
authors.forename: Blessing E
authors.initials: BE
authors.lastname: Obinaju
authors.affiliation: Division of Biomedical and Life Sciences, Faculty of Health and Medicine, Lancaster University, UK.
authors.forename: Nigel J
authors.initials: NJ
authors.lastname: Fullwood
authors.affiliation: Centre for Biophotonics, Lancaster Environment Centre, Lancaster University, Bailrigg, Lancaster LA1 4YQ, UK. Electronic address: f.martin@lancaster.ac.uk.
authors.forename: Francis L
authors.initials: FL
authors.lastname: Martin
mesh_terms: D001564:Benzo(a)pyrene / Q000633:toxicity*; D015415:Biomarkers / Q000378:metabolism; D001943:Breast Neoplasms / Q000378:metabolism / Q000648:ultrastructure*; D002467:Cell Nucleus / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D004249:DNA Damage; D005260:Female; D006801:Humans; D061986:MCF-7 Cells; D008855:Microscopy, Electron, Scanning; D015999:Multivariate Analysis; D016192:Resting Phase, Cell Cycle; D016196:S Phase; D017550:Spectroscopy, Fourier Transform Infrared; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D015415:Biomarkers; D001564:Benzo(a)pyrene
keywords: ATR-FTIR spectroscopy; Benzo[a]pyrene; DNA damage; MCF-7 cells; Nucleus isolation; Polycyclic aromatic hydrocarbon