Abstract
For many years, scientists have been looking for specific biomarkers associated with cancer cells for diagnosis purposes. These biomarkers mainly consist of proteins located at the cell surface (e.g. the TrkB receptor) whose activation is associated with specific metabolic modifications. Identification of these metabolic changes usually requires cell fixation and specific dye staining. MCARS microspectroscopy is a label-free, non-toxic, and minimally invasive method allowing to perform analyses of live cells and tissues. We used this method to follow the formation of lipid droplets in three colorectal cancer cell lines expressing TrkB. MCARS images of cells generated from signal integration of CH2 stretching modes allow to discriminate between lipid accumulation in the endoplasmic reticulum and the formation of cytoplasmic lipid droplets. We found that the number of the latter was related to the TrkB expression level. This result was confirmed thanks to the creation of a HEK cell line which over-expresses TrkB. We demonstrated that BDNF-induced TrkB activation leads to the formation of cytoplasmic lipid droplets, which can be abolished by K252a, an inhibitor of TrkB. So, MCARS microspectroscopy proved useful in characterizing cancer cells displaying an aberrant lipid metabolism.
Highlights
For many years, scientists have been looking for specific biomarkers associated with cancer cells for diagnosis purposes
The aim of our study was to use multiplex coherent anti-Stokes Raman scattering (MCARS) microspectroscopy in order to visualize and evaluate modifications of lipid metabolism induced by brain-derived neurotrophic factor (BDNF) binding to the TrkB receptor in TrkB-transfected HEK cells
The presence of the TrkB receptor was detected in three colorectal cancer cell lines representative of three different stages of the pathology: HCT116, HT29 and SW620
Summary
Scientists have been looking for specific biomarkers associated with cancer cells for diagnosis purposes These biomarkers mainly consist of proteins located at the cell surface (e.g. the TrkB receptor) whose activation is associated with specific metabolic modifications. MCARS microspectroscopy is a label-free, non-toxic, and minimally invasive method allowing to perform analyses of live cells and tissues We used this method to follow the formation of lipid droplets in three colorectal cancer cell lines expressing TrkB. The neurotrophin family is composed of three major receptors, Trk A, B and C, associated with specific ligands: NGF binds to TrkA, BDNF and NT-4 bind to TrkB and NT-3 binds to TrkC6 When they are expressed in other human tissues, neurotrophins are associated with tumorigenesis, metastases, for example in ovarian, breast, and colorectal c ancers[7]. MCARS microspectroscopy is a highly sensitive technology that is able, in the mid-infrared spectral domain, to visualize proteins, lipids and heterochromatin inside the c ells[9,10]
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