Combined Fourier transform infrared and Raman spectroscopic approach for identification of multidrug resistance phenotype in cancer cell lines

Abstract

Cancer cells escape cytotoxic effects of anticancer drugs by a process known as multidrug resistance (MDR). Identification of cell status by less time-consuming methods can be extremely useful in patient management and treatment. This study aims at evaluating the potentials of vibrational spectroscopic methods to perform cell typing and to differentiate between sensitive and resistant human cancer cell lines, in particular those that exhibit the MDR phenotype. Micro-Raman and Fourier transform infrared (FTIR) spectra have been acquired from the sensitive promyelocytic HL60 leukemia cell line and two of its subclones resistant to doxorubicin (HL60/DOX) and daunorubicin (HL60/ DNR), and from the sensitive MCF7 breast cancer cell line and its MDR counterpart resistant to verapamil (MCF7/VP). Principal components analysis (PCA) was employed for spectral comparison and classification. Our data show that cell typing was feasible with both methods, giving two distinct clusters for HL60-and MCF7-sensitive cells. In addition, phenotyping of HL60 cells, i.e., discriminating between the sensitive and MDR phenotypes, was attempted by both methods. FTIR could not only delineate between the sensitive and resistant HL60 cells, but also gave two distinct clusters for the resistant cells, which required a two-step procedure with Raman spectra. In the case of MCF7 cell lines, both the sensitive and resistant phenotypes could be differentiated very efficiently by PCA analysis of their FTIR and Raman point spectra. These results indicate the prospective applicability of FTIR and micro-Raman approaches in the differentiation of cell types as well as characterization of the cell status, such as the MDR phenotype exhibited in resistant leukemia cell lines like HL60 and MCF7.

Evaluation of the potential of Raman microspectroscopy for prediction of chemotherapeutic response to cisplatin in lung adenocarcinoma

“…Briefly, quartz coverslips were coated with a 2% w/v gelatin-water solution and maintained at 4 °C for 6 hrs to allow 65 polymerization of the gelatin to the substrate. Subsequently, 2.5  10 3 A549 cells were attached to the substrates for a 48 hour period, and were exposed to the cisplatin concentrations employed for the cytotoxicity assay for a 96-hour period (together with a non-exposed 70 control sample). After the exposure period, the cells were fixed in 4% formalin for 10 minutes and were stored in 0.9 % physiological saline solution at 4 °C until the Raman analysis was performed.…”

“…The aim of the current study is to evaluate the quantitative 70 predictive capabilities of CRM in the elucidation of the mode of action of chemotherapeutic agents, using cisplatin and A549 adenocarcinoma cells as a model compound and test system respectively. Spectra of the cellular nuclei were taken after a 96 hour exposure period to the agent, and 75 multivariate models of the variation in spectral content with levels of exposure and with degrees of cytotoxicological response were constructed.…”

Application of Fourier Transform Infrared Spectroscopy for Tumor Diagnosis

“…Recently some spectral methods have been applied for evaluation of malignancies, and some attempts have been made to utilize some of these tests as screening techniques (4,11,31,35,36,43). infrared spectroscopy is a powerful tool for studies of the composition and structure of cellular components (1,49,50).…”

Renal Graft Fibrosis and Inflammation Quantification by an Automated Fourier–Transform Infrared Imaging Technique

“…This technique has proven effective to discriminate between tumor and normal tissues 14,15 and between benign and malignant lesions 14,16 and also, identify drug resistance in cancer. 17,18 Infrared spectroscopy has also been used to identify renal stones 19,20 and characterize renal tumor tissues. 21 Nonetheless, to the best of our knowledge, no study has been published to date showing the possible contribution of infrared imaging to the characterization of nontumor renal tissue, particularly in terms of IF and inflammation.…”