title: Discrimination of bladder cancer cells from normal urothelial cells with high specificity and sensitivity: combined application of atomic force microscopy and modulated Raman spectroscopy.
abstract: Atomic force microscopy (AFM) and modulated Raman spectroscopy (MRS) were used to discriminate between living normal human urothelial cells (SV-HUC-1) and bladder tumour cells (MGH-U1) with high specificity and sensitivity. MGH-U1 cells were 1.5-fold smaller, 1.7-fold thicker and 1.4-fold rougher than normal SV-HUC-1 cells. The adhesion energy was 2.6-fold higher in the MGH-U1 cells compared to normal SV-HUC-1 cells, which possibly indicates that bladder tumour cells are more deformable than normal cells. The elastic modulus of MGH-U1 cells was 12-fold lower than SV-HUC-1 cells, suggesting a higher elasticity of the bladder cancer cell membranes. The biochemical fingerprints of cancer cells displayed a higher DNA and lipid content, probably due to an increase in the nuclear to cytoplasm ratio. Normal cells were characterized by higher protein contents. AFM studies revealed a decrease in the lateral dimensions and an increase in thickness of cancer cells compared to normal cells; these studies authenticate the observations from MRS. Nanostructural, nanomechanical and biochemical profiles of bladder cells provide qualitative and quantitative markers to differentiate between normal and cancerous cells at the single cellular level. AFM and MRS allow discrimination between adhesion energy, elasticity and Raman spectra of SV-HUC-1 and MGH-U1 cells with high specificity (83, 98 and 95%) and sensitivity (97, 93 and 98%). Such single-cell-level studies could have a pivotal impact on the development of AFM-Raman combined methodologies for cancer profiling and screening with translational significance.
authors.affiliation: School of Science, Engineering and Technology, Abertay University, Bell Street, Dundee DD1 1HG, UK; School of Physics and Astronomy, University of St Andrews, North Haugh, Fife KY16 9SS, UK.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Canetta
authors.affiliation: School of Medicine, University of St Andrews, St Andrews, Fife KY16 9TF, UK.
authors.forename: Andrew
authors.initials: A
authors.lastname: Riches
authors.affiliation: School of Biology, University of St Andrews, St Andrews, Fife KY16 9TF, UK.
authors.forename: Eva
authors.initials: E
authors.lastname: Borger
authors.affiliation: School of Medicine, University of St Andrews, St Andrews, Fife KY16 9TF, UK.
authors.forename: Simon
authors.initials: S
authors.lastname: Herrington
authors.affiliation: School of Physics and Astronomy, University of St Andrews, North Haugh, Fife KY16 9SS, UK.
authors.forename: Kishan
authors.initials: K
authors.lastname: Dholakia
authors.affiliation: School of Science, Engineering and Technology, Abertay University, Bell Street, Dundee DD1 1HG, UK. Electronic address: ashok.adya@abertay.ac.uk.
authors.forename: Ashok K
authors.initials: AK
authors.lastname: Adya
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000473:pathology; D055119:Elastic Modulus; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods*; D049329:Nanostructures / Q000648:ultrastructure; D010590:Phalloidine / Q000378:metabolism; D025341:Principal Component Analysis; D012680:Sensitivity and Specificity; D013059:Spectrum Analysis, Raman / Q000379:methods*; D001749:Urinary Bladder Neoplasms / Q000473:pathology*; D019459:Urothelium / Q000473:pathology*
chemical_list: D000199:Actins; D010590:Phalloidine
keywords: Atomic force microscopy; Bladder cancer; Cell mechanics; Cytoskeleton organization; Modulated Raman spectroscopy