title: Nanoscale characterization illustrates the cisplatin-mediated biomechanical changes of B16-F10 melanoma cells.
abstract: Cells reorganize their membrane biomechanical dynamics in response to environmental stimuli or inhibitors associated with their physiological/pathological processes, and disease therapeutics. To validate the biophysical dynamics during cell exposure to anti-cancer drugs, we investigate the nanoscale biological characterization in melanoma cells undergoing cisplatin treatment. Using atomic force microscopy, we demonstrate that the cellular morphology and membrane ultrastructure are altered after exposure to cisplatin. In contrast to their normal spindle-like shape, cisplatin causes cell deformation rendering cells flat and enlarged, which increases the cell area by 3-4 fold. Additionally, cisplatin decreases the topography height values for both the cytoplasmic and nuclear regions (by 40-80% and 60%, respectively). Furthermore, cisplatin increases the cytoplasmic root mean square roughness by 110-240% in correlation with the drug concentration and attenuates the nuclear RMS by 60%. Moreover, the cellular adhesion force was enhanced, while the Young's modulus elasticity was attenuated by ∼2 and ∼2.3 fold, respectively. F-actin phalloidin staining revealed that cisplatin enlarges the cell size through enhanced stress fiber formation and promotes cytoskeletal reorganization. Immunoblot analyses further revealed that the activities of focal adhesion proteins, such as FAK and c-Src, are upregulated by cisplatin through phosphorylation at tyrosine 397 and 530, respectively. Collectively, these results show that cisplatin-treated melanoma cells not only exhibit the upregulation of FAK-mediated signaling to enhance the cytoskeleton mechanical stretch, but also promote the cytoskeletal rearrangement resulting in 43% decrease in the cell modulus. These mechanisms thus promote the malignancy and invasiveness of the melanoma cells.
authors.affiliation: Department of Chemistry, National Sun Yat-sen University, 70 Lien-hai Rd., Kaohsiung 80424, Taiwan. shsieh@faculty.nsysu.edu.tw.
authors.forename: Mei-Lang
authors.initials: ML
authors.lastname: Kung
authors.affiliation: Department of Chemistry, National Sun Yat-sen University, 70 Lien-hai Rd., Kaohsiung 80424, Taiwan. shsieh@faculty.nsysu.edu.tw.
authors.forename: Chiung-Wen
authors.initials: CW
authors.lastname: Hsieh
authors.affiliation: Department of Biological Sciences, National Sun Yat-Sen University, Kaohsiung 80424, Taiwan and Institute of Biomedical Sciences, National Sun Yat-sen University, Kaohsiung 80424, Taiwan and Center for Neuroscience, National Sun Yat-Sen University, Kaohsiung 80424, Taiwan and Center for Stem Cell Research, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
authors.forename: Ming-Hong
authors.initials: MH
authors.lastname: Tai
authors.affiliation: Department of Biological Sciences, National Sun Yat-Sen University, Kaohsiung 80424, Taiwan.
authors.forename: Chien-Hui
authors.initials: CH
authors.lastname: Weng
authors.affiliation: Center for Stem Cell Research, Kaohsiung Medical University, Kaohsiung 80708, Taiwan and Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung 80708, Taiwan and Department of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan and Department of Internal Medicine, Kaohsiung Municipal Ta-Tung Hospital, Kaohsiung 80708, Taiwan.
authors.forename: Deng-Chyang
authors.initials: DC
authors.lastname: Wu
authors.affiliation: Center for Stem Cell Research, Kaohsiung Medical University, Kaohsiung 80708, Taiwan and Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan and Department of Urology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung 80708, Taiwan and Department of Urology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan and Department of Urology, Kaohsiung Municipal Ta-Tung Hospital, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
authors.forename: Wen-Jeng
authors.initials: WJ
authors.lastname: Wu
authors.affiliation: Center for Stem Cell Research, Kaohsiung Medical University, Kaohsiung 80708, Taiwan and Department of Urology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung 80708, Taiwan and Department of Urology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
authors.forename: Bi-Wen
authors.initials: BW
authors.lastname: Yeh
authors.affiliation: Department of Seafood Science, National Kaohsiung Marine University, Kaohsiung 80811, Taiwan.
authors.forename: Shu-Ling
authors.initials: SL
authors.lastname: Hsieh
authors.affiliation: Center for Stem Cell Research, Kaohsiung Medical University, Kaohsiung 80708, Taiwan and Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung 80708, Taiwan and Department of Internal Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
authors.forename: Chao-Hung
authors.initials: CH
authors.lastname: Kuo
authors.affiliation: Graduate Institute of Basic Medical Science, China Medical University, Taichung 40402, Taiwan and Center for Neuropsychiatry, China Medical University Hospital, Taichung 40402, Taiwan.
authors.forename: Huey-Shan
authors.initials: HS
authors.lastname: Hung
authors.affiliation: Department of Chemistry, National Sun Yat-sen University, 70 Lien-hai Rd., Kaohsiung 80424, Taiwan. shsieh@faculty.nsysu.edu.tw and Center for Stem Cell Research, Kaohsiung Medical University, Kaohsiung 80708, Taiwan and School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
authors.forename: Shuchen
authors.initials: S
authors.lastname: Hsieh
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001696:Biomechanical Phenomena; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D048429:Cell Size; D002945:Cisplatin / Q000494:pharmacology*; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D008546:Melanoma, Experimental / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D018625:Microscopy, Atomic Force; D036103:Nanotechnology; D015398:Signal Transduction; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D051416:Focal Adhesion Protein-Tyrosine Kinases; D019061:src-Family Kinases; D002945:Cisplatin