In situ Fourier transform infrared analysis of live cells' response to doxorubicin

TY - JOUR

T1 - In situ Fourier transform infrared analysis of live cells' response to doxorubicin

AU - Vieira Fale, Pedro

AU - Altharawi, Ali

AU - Chan, K. L Andrew

PY - 2015/10/1

Y1 - 2015/10/1

N2 - The study of the response of cancer cells to chemotherapy drugs is of high importance due to the specificity of some drugs to certain types of cancer and the resistance of some specific cancer types to chemotherapy drugs. Our aim was to develop and apply the label-free and non-destructive Fourier transform infrared (FTIR) method to determine the sensitivity of three different cancer cell-lines to a common anti-cancer drug doxorubicin at different concentrations and to demonstrate that information about the mechanism of resistance to the chemotherapy drug can be extracted from spectral data. HeLa, PC3, and Caco-2 cells were seeded and grown on an attenuated total reflection (ATR) crystal, doxorubicin was applied at the clinically significant concentration of 0.1-20μM, and spectra of the cells were collected hourly over 20h. Analysis of the amide bands was correlated with cell viability, which had been cross validated with MTT assays, allowing to determine that the three cell lines had significantly different resistance to doxorubicin. The difference spectra and principal component analysis (PCA) highlighted the subtle chemical changes in the living cells under treatment. Spectral regions assigned to nucleic acids (mainly 1085cm-1) and carbohydrates (mainly 1024cm-1) showed changes that could be related to the mode of action of the drug and the mechanism of resistance of the cell lines to doxorubicin. This is a cost-effective method that does not require bioassay reagents but allows label-free, non-destructive and in situ analysis of chemical changes in live cells, using standard FTIR equipment adapted to ATR measurements.

AB - The study of the response of cancer cells to chemotherapy drugs is of high importance due to the specificity of some drugs to certain types of cancer and the resistance of some specific cancer types to chemotherapy drugs. Our aim was to develop and apply the label-free and non-destructive Fourier transform infrared (FTIR) method to determine the sensitivity of three different cancer cell-lines to a common anti-cancer drug doxorubicin at different concentrations and to demonstrate that information about the mechanism of resistance to the chemotherapy drug can be extracted from spectral data. HeLa, PC3, and Caco-2 cells were seeded and grown on an attenuated total reflection (ATR) crystal, doxorubicin was applied at the clinically significant concentration of 0.1-20μM, and spectra of the cells were collected hourly over 20h. Analysis of the amide bands was correlated with cell viability, which had been cross validated with MTT assays, allowing to determine that the three cell lines had significantly different resistance to doxorubicin. The difference spectra and principal component analysis (PCA) highlighted the subtle chemical changes in the living cells under treatment. Spectral regions assigned to nucleic acids (mainly 1085cm-1) and carbohydrates (mainly 1024cm-1) showed changes that could be related to the mode of action of the drug and the mechanism of resistance of the cell lines to doxorubicin. This is a cost-effective method that does not require bioassay reagents but allows label-free, non-destructive and in situ analysis of chemical changes in live cells, using standard FTIR equipment adapted to ATR measurements.

KW - Chemotherapy

KW - Drug resistance

KW - Drug screening

KW - Infrared spectroscopy

KW - Living cells

KW - Multi-bounce ATR

UR - http://www.scopus.com/inward/record.url?scp=84938814184&partnerID=8YFLogxK

U2 - 10.1016/j.bbamcr.2015.07.018

DO - 10.1016/j.bbamcr.2015.07.018

M3 - Article

AN - SCOPUS:84938814184

SN - 0167-4889

VL - 1853

SP - 2640

EP - 2648

JO - Biochimica et biophysica acta-Molecular cell research

JF - Biochimica et biophysica acta-Molecular cell research

IS - 10 Part A

ER -