title: Different Phases of Breast Cancer Cells: Raman Study of Immortalized, Transformed, and Invasive Cells.
abstract: Breast cancer is the most prevalent cause of cancer-associated death in women the world over, but if detected early it can be treated successfully. Therefore, it is important to diagnose this disease at an early stage and to understand the biochemical changes associated with cellular transformation and cancer progression. Deregulated lipid metabolism has been shown to contribute to cell transformation as well as cancer progression. In this study, we monitored the biomolecular changes associated with the transformation of a normal cell into an invasive cell associated with breast cancer using Raman microspectroscopy. We have utilized primary normal breast cells, and immortalized, transformed, non-invasive, and invasive breast cancer cells. The Raman spectra were acquired from all these cell lines under physiological conditions. The higher wavenumber (2800-3000 cm-1) and lower wavenumber (700-1800 cm-1) range of the Raman spectrum were analyzed and we observed increased lipid levels for invasive cells. The Raman spectral data were analyzed by principal component-linear discriminant analysis (PC-LDA), which resulted in the formation of distinct clusters for different cell types with a high degree of sensitivity. The subsequent testing of the PC-LDA analysis via the leave-one-out cross validation approach (LOOCV) yielded relatively high identification sensitivity. Additionally, the Raman spectroscopic results were confirmed through fluorescence staining tests with BODIPY and Nile Red biochemical assays. Furthermore, Raman maps from the above mentioned cells under fixed conditions were also acquired to visualize the distribution of biomolecules throughout the cell. The present study shows the suitability of Raman spectroscopy as a non-invasive, label-free, microspectroscopic technique, having the potential of probing changes in the biomolecular composition of living cells as well as fixed cells.
authors.affiliation: Department of Inorganic and Physical Chemistry, Indian Institute of Science, Bangalore 560012, India. deepika.5185@gmail.com.
authors.forename: Deepika
authors.initials: D
authors.lastname: Chaturvedi
authors.affiliation: Molecular Reproduction, Development & Genetics, Indian Institute of Science, Bangalore 560012, India. balu.1pharma@gmail.com.|Department of Pharmacology, Manipal College of Pharmaceutical sciences, Manipal 576104, India. balu.1pharma@gmail.com.
authors.forename: Sai A
authors.initials: SA
authors.lastname: Balaji
authors.affiliation: Department of Inorganic and Physical Chemistry, Indian Institute of Science, Bangalore 560012, India. vinaykbn07@gmail.com.
authors.forename: Vinay Kumar
authors.initials: VK
authors.lastname: Bn
authors.affiliation: Department of Inorganic and Physical Chemistry, Indian Institute of Science, Bangalore 560012, India. f.ariese@vu.nl.
authors.forename: Freek
authors.initials: F
authors.lastname: Ariese
authors.affiliation: Department of Inorganic and Physical Chemistry, Indian Institute of Science, Bangalore 560012, India. umapathy@ipc.iisc.ernet.in.|Department of Instrumentation and Applied Physics, Indian Institute of Science, Bangalore 560012, India. umapathy@ipc.iisc.ernet.in.
authors.forename: Siva
authors.initials: S
authors.lastname: Umapathy
authors.affiliation: Molecular Reproduction, Development & Genetics, Indian Institute of Science, Bangalore 560012, India. umapathy_india@yahoo.com.
authors.forename: Annapoorni
authors.initials: A
authors.lastname: Rangarajan
mesh_terms: D001943:Breast Neoplasms / Q000175:diagnosis / Q000378:metabolism* / Q000473:pathology; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000378:metabolism; D016002:Discriminant Analysis; D005260:Female; D006801:Humans; D050356:Lipid Metabolism; D013059:Spectrum Analysis, Raman
keywords: LOOCV; PC-LDA; Raman spectroscopy; breast cancer; lipid staining