Identification of morphological and biochemical changes in keratin-8/18 knock-down cells using Raman spectroscopy

5 years ago

Identification of morphological and biochemical changes in keratin-8/18 knock-down cells using Raman spectroscopy

S. P. Singh, Crismita Dmello, C. Murali Krishna, Milind M. Vaidya, Hitesh Mamgain, Hunain Alam, Ramachandra Rao Dasari

Accurate understanding of cellular processes and responses to stimuli is of paramount importance in biomedical research and diagnosis. Raman spectroscopy (RS), a label-free and nondestructive spectroscopic method has the potential to serve as a novel ’theranostics' tool. Both fiber-optic and micro-Raman studies have demonstrated efficacy in diagnostics and therapeutic response monitoring. In the present study, we have evaluated the potential of micro-Raman spectroscopic maps in identifying changes induced by loss of K8/18 proteins in a tongue cancer cell line. Furthermore, we also evaluated the efficacy of less expensive and commercially available fiber probes to identify K8/18 wild and knock-down cell pellets, in view of the utility of cell pellet-based studies. The findings suggest that major differences in the cellular morphology and biochemical composition can be objectively identified and can be utilized for classification using both micro-Raman and fiber-probe-based RS. These findings highlight the potential of fiber-optic probe-based RS in noninvasive cellular phenotyping for diagnosis and therapeutic response monitoring, especially in low-resource settings. In the present study, Raman spectroscopy, both fiber-optic and microscopy, were explored to study changes due to loss of ‘keratin' protein in tongue cancer cells. A microscopic approach was explored to study changes at the single-cell level and applicability of less expensive and commercially available for fiber-optic probes on cell pellets was also assessed. The findings can help in designing novel keratin-based ‘theranostics' tools for oral cancers.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/jbio.201600249