title: Stiffness changes of tumor HEp2 cells correlates with the inhibition and release of TRAIL-induced apoptosis pathways.
abstract: Tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL) is a promising apoptotic agent that can selectively act on tumor cells. However, some cancer cells are resistant to TRAIL mediated apoptosis. In specific type of cells, sensitization by chemotherapeutic drugs may overcome the resistance to TRAIL induced apoptosis. In this work, atomic force microscopy (AFM) nanoindentation spectroscopy combined with fluorescence methods were used to investigate the biomechanical aspects of the resistance and unblocking of apoptosis in larynx carcinoma HEp2 cells treated with TRAIL. It is shown that there is a direct correlation between the increase in mechanical cell stiffness and the inhibition of apoptosis induced by TRAIL in HEp2 cells. Conversely, unblocking of apoptosis by sensitization of HEp2 cells with a chemotherapeutic drug Actinomycin D is related to the depolymerization of F-actin and to the decrease in the cell stiffness. Both effects, that is, changes in the mechanical stiffness of the cell and the inhibition of apoptotic pathway, are closely related to the Bcl-2 activity. Most probably, the depolymerization of F-actin results from downregulation of Rho protein, which in turn is accompanied by a lower activity of Bcl-2 and in consequence releases the intrinsic apoptotic channel. The presented results reveal a promising application of nanoindentation spectroscopy with an AFM tip as a novel tool for monitoring the processes of apoptosis inhibition.
authors.affiliation: Institute of Physics, Jagiellonian University, Reymonta 4, 30-059 Krakow, Poland. marta.targosz-korecka@uj.edu.pl
authors.forename: Marta
authors.initials: M
authors.lastname: Targosz-Korecka
authors.forename: Rafal
authors.initials: R
authors.lastname: Biedron
authors.forename: Agnieszka Maria
authors.initials: AM
authors.lastname: Szczygiel
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Brzezinka
authors.forename: Jacek
authors.initials: J
authors.lastname: Szczerbinski
authors.forename: Anna
authors.initials: A
authors.lastname: Zuk
mesh_terms: D000199:Actins / Q000378:metabolism; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D017209:Apoptosis; D003609:Dactinomycin / Q000494:pharmacology; D004359:Drug Therapy, Combination; D004548:Elasticity; D005453:Fluorescence; D006801:Humans; D007822:Laryngeal Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D018625:Microscopy, Atomic Force; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
chemical_list: D000199:Actins; D000903:Antibiotics, Antineoplastic; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; D003609:Dactinomycin