TY - JOUR

T1 - Identification of benzo[a]pyrene-induced cell cycle-associated alterations in MCF-7 cells using infrared spectroscopy with computational analysis

AU - Pang, Weiyi

AU - Li, Junyi

AU - Ahmadzai, Abdullah A

AU - Heppenstall, Lara D

AU - Llabjani, Valon

AU - Trevisan, Júlio

AU - Qiu, Xiaoqiang

AU - Martin, Francis L

PY - 2012/8/16

Y1 - 2012/8/16

N2 - Chemical contaminants, such as benzo[a]pyrene (B[a]P), may modulate transcriptional responses in cells via the activation of aryl hydrocarbon receptor (AhR) or through responses to DNA damage following adduct formation. Attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy can be employed in a non-destructive fashion to interrogate the biochemical signature of cells via generation of infrared (IR) spectra. By applying to generated spectral datasets subsequent computational approaches such as principal component analysis plus linear discriminant analysis (PCA-LDA), derived data reduction is achieved to facilitate the visualization of wavenumber-related alterations in target cells. Discriminating spectral variables might be associated with lipid or glycogen content, conformational protein changes and phosphorylation, and structural alterations in DNA/RNA. Using this approach, we investigated the dose-related effects of B[a]P in MCF-7 cells concentrated in S- or G₀/G₁-phase. Our findings identified that in PCA-LDA scores plots a clear segregation of IR spectra was evident, with the major spectral alterations associated with DNA/RNA, secondary protein structure and lipid. Dose-related effects were observed and even with exposures as low as 10⁻⁹ M B[a]P, significant (P ≤ 0.001) separation of B[a]P-treated vs. vehicle control cells was noted. ATR-FTIR spectroscopy with computational analysis is a novel approach to identify the effects of environmental contaminants in target cells.

AB - Chemical contaminants, such as benzo[a]pyrene (B[a]P), may modulate transcriptional responses in cells via the activation of aryl hydrocarbon receptor (AhR) or through responses to DNA damage following adduct formation. Attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy can be employed in a non-destructive fashion to interrogate the biochemical signature of cells via generation of infrared (IR) spectra. By applying to generated spectral datasets subsequent computational approaches such as principal component analysis plus linear discriminant analysis (PCA-LDA), derived data reduction is achieved to facilitate the visualization of wavenumber-related alterations in target cells. Discriminating spectral variables might be associated with lipid or glycogen content, conformational protein changes and phosphorylation, and structural alterations in DNA/RNA. Using this approach, we investigated the dose-related effects of B[a]P in MCF-7 cells concentrated in S- or G₀/G₁-phase. Our findings identified that in PCA-LDA scores plots a clear segregation of IR spectra was evident, with the major spectral alterations associated with DNA/RNA, secondary protein structure and lipid. Dose-related effects were observed and even with exposures as low as 10⁻⁹ M B[a]P, significant (P ≤ 0.001) separation of B[a]P-treated vs. vehicle control cells was noted. ATR-FTIR spectroscopy with computational analysis is a novel approach to identify the effects of environmental contaminants in target cells.

KW - ATR-FTIR spectroscopy

KW - Benzo[a]pyrene

KW - Cell cycle

KW - Computational analysis

KW - MCF-7 cells

KW - PCA–LDA

U2 - 10.1016/j.tox.2012.04.009

DO - 10.1016/j.tox.2012.04.009

M3 - Journal article

C2 - 22561278

VL - 298

SP - 24

EP - 29

JO - Toxicology

JF - Toxicology

IS - 1-3

ER -