{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,8]],"date-time":"2026-02-08T08:58:00Z","timestamp":1770541080513,"version":"3.49.0"},"reference-count":46,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2022,10,15]],"date-time":"2022-10-15T00:00:00Z","timestamp":1665792000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"FCT","award":["LA\/P\/0070\/2020"],"award-info":[{"award-number":["LA\/P\/0070\/2020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Colorectal cancer (CRC) has been addressed in the framework of molecular, cellular biology, and biochemical traits. A new approach to studying CRC is focused on the relationship between biochemical pathways and biophysical cues, which may contribute to disease understanding and therapy development. Herein, we investigated the mechanical properties of CRC cells, namely, HCT116, HCT15, and SW620, using static and dynamic methodologies by atomic force microscopy (AFM). The static method quantifies Young\u2019s modulus; the dynamic method allows the determination of elasticity, viscosity, and fluidity. AFM results were correlated with confocal laser scanning microscopy and cell migration assay data. The SW620 metastatic cells presented the highest Young\u2019s and storage moduli, with a defined cortical actin ring with distributed F-actin filaments, scarce vinculin expression, abundant total focal adhesions (FAK), and no filopodia formation, which could explain the lessened migratory behavior. In contrast, HCT15 cells presented lower Young\u2019s and storage moduli, high cortical tubulin, less cortical F-actin and less FAK, and more filopodia formation, probably explaining the higher migratory behavior. HCT116 cells presented Young\u2019s and storage moduli values in between the other cell lines, high cortical F-actin expression, intermediate levels of total FAK, and abundant filopodia formation, possibly explaining the highest migratory behavior.<\/jats:p>","DOI":"10.3390\/cancers14205053","type":"journal-article","created":{"date-parts":[[2022,10,16]],"date-time":"2022-10-16T21:10:10Z","timestamp":1665954610000},"page":"5053","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Mechanical Properties of Colorectal Cancer Cells Determined by Dynamic Atomic Force Microscopy: A Novel Biomarker"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6638-1211","authenticated-orcid":false,"given":"M. Manuela","family":"Br\u00e1s","sequence":"first","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade (i3S), Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Instituto de Engenharia Biom\u00e9dica (INEB), Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Faculdade de Engenharia da Universidade do Porto, 4200-465 Porto, Portugal"}]},{"given":"T\u00e2nia B.","family":"Cruz","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade (i3S), Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Instituto de Engenharia Biom\u00e9dica (INEB), Universidade do Porto, 4200-135 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5987-7016","authenticated-orcid":false,"given":"Andr\u00e9 F.","family":"Maia","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade (i3S), Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Instituto de Biologia Molecular e Celular (IBMC), Universidade do Porto, 4200-135 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0724-0272","authenticated-orcid":false,"given":"Maria Jos\u00e9","family":"Oliveira","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade (i3S), Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Instituto de Engenharia Biom\u00e9dica (INEB), Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Instituto de Ci\u00eancias Biom\u00e9dicas Abel Salazar (ICBAS), Universidade do Porto, 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8018-4310","authenticated-orcid":false,"given":"Susana R.","family":"Sousa","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade (i3S), Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Instituto de Engenharia Biom\u00e9dica (INEB), Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Instituto Superior de Engenharia do Porto (ISEP), Instituto Polit\u00e9cnico do Porto, 4200-072 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2761-4929","authenticated-orcid":false,"given":"Pedro L.","family":"Granja","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade (i3S), Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Instituto de Engenharia Biom\u00e9dica (INEB), Universidade do Porto, 4200-135 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8744-4541","authenticated-orcid":false,"given":"Manfred","family":"Radmacher","sequence":"additional","affiliation":[{"name":"Institute of Biophysics, University of Bremen, 28334 Bremen, Germany"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"209","DOI":"10.3322\/caac.21660","article-title":"Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA A Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"104282","DOI":"10.1016\/j.phrs.2019.104282","article-title":"Targeting Ca2+ signalling through phytomolecules to combat cancer","volume":"146","author":"Singh","year":"2019","journal-title":"Pharmacol. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1093\/gastro\/got041","article-title":"Colorectal polyps and polyposis syndromes","volume":"2","author":"Shussman","year":"2014","journal-title":"Gastroenterol. Rep."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"871","DOI":"10.1016\/j.cell.2009.11.007","article-title":"Epithelial-mesenchymal transitions in development and disease","volume":"139","author":"Thiery","year":"2009","journal-title":"Cell"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1038\/nrc.2018.15","article-title":"Cell motility in cancer invasion and metastasis: Insights from simple model organisms","volume":"18","author":"Stuelten","year":"2018","journal-title":"Nat. Rev. Cancer"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1537","DOI":"10.1016\/j.bbagen.2018.02.009","article-title":"Transforming growth factor-\u03b2 modulates pancreatic cancer associated fibroblasts cell shape, stiffness and invasion","volume":"1862","author":"Stylianou","year":"2018","journal-title":"Biochim. Biophys. Acta (BBA)-Gen. Subj."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1093","DOI":"10.1016\/j.ymthe.2018.01.021","article-title":"Autologous Cell Therapy Approach for Duchenne Muscular Dystrophy using PiggyBac Transposons and Mesoangioblasts","volume":"26","author":"Iyer","year":"2018","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"5620","DOI":"10.1021\/acsbiomaterials.0c00975","article-title":"Tissue Rheology as a Possible Complementary Procedure to Advance Histological Diagnosis of Colon Cancer","volume":"6","author":"Pogoda","year":"2020","journal-title":"ACS Biomater. Sci. Eng."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"045703","DOI":"10.1088\/1361-6528\/28\/4\/045703","article-title":"Nanoindentation characterisation of human colorectal cancer cells considering cell geometry, surface roughness and hyperelastic constitutive behaviour","volume":"28","author":"Boccaccio","year":"2017","journal-title":"Nanotechnology"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1007\/s00424-008-0524-3","article-title":"Elasticity measurement of living cells with an atomic force microscope: Data acquisition and processing","volume":"457","author":"Carl","year":"2008","journal-title":"Pflug. Arch. Eur. J. Physiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"335401","DOI":"10.1088\/1361-6463\/ac02fa","article-title":"Multiple power-law viscoelastic relaxation in time and frequency domains with atomic force microscopy","volume":"54","author":"Babu","year":"2021","journal-title":"J. Phys. D Appl. Phys."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1007\/s10585-012-9531-z","article-title":"Cytoskeletal stiffness, friction, and fluidity of cancer cell lines with different metastatic potential","volume":"30","author":"Coughlin","year":"2013","journal-title":"Clin. Exp. Metastasis"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"54","DOI":"10.3389\/fcell.2020.00054","article-title":"Melanoma in the Eyes of Mechanobiology","volume":"8","author":"Radmacher","year":"2020","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"10771","DOI":"10.1038\/s41598-021-90173-y","article-title":"Cell mechanical properties of human breast carcinoma cells depend on temperature","volume":"11","author":"Aermes","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"021914","DOI":"10.1103\/PhysRevE.72.021914","article-title":"Probing mechanical properties of living cells by atomic force microscopy with blunted pyramidal cantilever tips","volume":"72","author":"Rico","year":"2005","journal-title":"Phys. Rev. E"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"5117","DOI":"10.1038\/s41598-017-05383-0","article-title":"Standardized Nanomechanical Atomic Force Microscopy Procedure (SNAP) for Measuring Soft and Biological Samples","volume":"7","author":"Schillers","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"716","DOI":"10.1021\/la0110850","article-title":"Correction of Microrheological Measurements of Soft Samples with Atomic Force Microscopy for the Hydrodynamic Drag on the Cantilever","volume":"18","author":"Alcaraz","year":"2002","journal-title":"Langmuir ACS J. Surf. Colloids"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.semcdb.2017.07.022","article-title":"The role of the microenvironment in the biophysics of cancer","volume":"Volume 73","author":"Rianna","year":"2018","journal-title":"Seminars in Cell & Developmental Biology"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2071","DOI":"10.1016\/S0006-3495(03)75014-0","article-title":"Microrheology of human lung epithelial cells measured by atomic force microscopy","volume":"84","author":"Alcaraz","year":"2003","journal-title":"Biophys. J."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Lin, C.Y., Chen, Y.C., Lin, C.H., and Chang, K.V. (2022). Constitutive Equations for Analyzing Stress Relaxation and Creep of Viscoelastic Materials Based on Standard Linear Solid Model Derived with Finite Loading Rate. Polymers, 14.","DOI":"10.3390\/polym14102124"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An open-source platform for biological-image analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"20130038","DOI":"10.1098\/rsfs.2013.0038","article-title":"Introduction to cell-hydrogel mechanosensing","volume":"4","author":"Ahearne","year":"2014","journal-title":"Interface Focus"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Ronen, J., Hayat, S., and Akalin, A. (2019). Evaluation of colorectal cancer subtypes and cell lines using deep learning. Life Sci. Alliance, 2.","DOI":"10.26508\/lsa.201900517"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.jare.2013.07.006","article-title":"Hydrogel: Preparation, characterization, and applications","volume":"6","author":"Ahmed","year":"2015","journal-title":"J. Adv. Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/j.micron.2009.10.008","article-title":"Effect of AEE788 and\/or Celecoxib on colon cancer cell morphology using advanced microscopic techniques","volume":"41","author":"Venkatesan","year":"2010","journal-title":"Micron"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2027079","DOI":"10.1155\/2017\/2027079","article-title":"Effects of Tumor Necrosis Factor-\u03b1 on Morphology and Mechanical Properties of HCT116 Human Colon Cancer Cells Investigated by Atomic Force Microscopy","volume":"2017","author":"Liu","year":"2017","journal-title":"Scanning"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1002\/jib.139","article-title":"Antiproliferative effects of prenylflavonoids from hops on human colon cancer cell lines","volume":"120","author":"Fiala","year":"2014","journal-title":"J. Inst. Brew."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4749","DOI":"10.1038\/s41598-020-61631-w","article-title":"Double power-law viscoelastic relaxation of living cells encodes motility trends","volume":"10","author":"Freire","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"472","DOI":"10.3389\/fphy.2021.711860","article-title":"Oscillatory Microrheology, Creep Compliance and Stress Relaxation of Biological Cells Reveal Strong Correlations as Probed by Atomic Force Microscopy","volume":"9","author":"Flormann","year":"2021","journal-title":"Front. Phys."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"5719","DOI":"10.1039\/C5SM01089F","article-title":"Mechanical and structural comparison between primary tumor and lymph node metastasis cells in colorectal cancer","volume":"11","author":"Palmieri","year":"2015","journal-title":"Soft Matter"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1038\/labinvest.2015.151","article-title":"Tyrosine 397 phosphorylation is critical for FAK-promoted Rac1 activation and invasive properties in oral squamous cell carcinoma cells","volume":"96","author":"Chiu","year":"2016","journal-title":"Lab. Investig."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"117","DOI":"10.2739\/kurumemedj.52.117","article-title":"Immunohistochemical distribution of tubulin beta II in human normal and neoplastic tissues","volume":"52","author":"Oda","year":"2005","journal-title":"Kurume Med. J."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Ruksha, K., Mezheyeuski, A., Nerovnya, A., Bich, T., Tur, G., Gorgun, J., Luduena, R., and Portyanko, A. (2019). Over-Expression of \u03b2II-Tubulin and Especially Its Localization in Cell Nuclei Correlates with Poorer Outcomes in Colorectal Cancer. Cells, 8.","DOI":"10.3390\/cells8010025"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"968","DOI":"10.1016\/j.semcdb.2011.09.017","article-title":"Regulation of cell migration by dynamic microtubules","volume":"22","year":"2011","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"532","DOI":"10.1007\/s12195-014-0345-8","article-title":"Substrate Stiffness Mediated Metastasis Like Phenotype of Colon Cancer Cells is Independent of Cell to Gel Adhesion","volume":"7","author":"Ali","year":"2014","journal-title":"Cell. Mol. Bioeng."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1016\/j.jmps.2013.08.018","article-title":"Probing mechanical principles of cell-nanomaterial interactions","volume":"62","author":"Gao","year":"2014","journal-title":"J. Mech. Phys. Solids"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"6766","DOI":"10.1073\/pnas.1820567116","article-title":"Structural and mechanistic insights into mechanoactivation of focal adhesion kinase","volume":"116","author":"Bauer","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Bao, Y., Guo, Y., Zhang, C., Fan, F., and Yang, W. (2017). Sphingosine Kinase 1 and Sphingosine-1-Phosphate Signaling in Colorectal Cancer. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18102109"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1043","DOI":"10.1083\/jcb.200703036","article-title":"Vinculin controls focal adhesion formation by direct interactions with talin and actin","volume":"179","author":"Humphries","year":"2007","journal-title":"J. Cell Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1038\/s12276-020-0389-x","article-title":"New role of the antidepressant imipramine as a Fascin1 inhibitor in colorectal cancer cells","volume":"52","author":"Rodrigues","year":"2020","journal-title":"Exp. Mol. Med."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"23420","DOI":"10.1074\/jbc.M110.102830","article-title":"Vinculin is a dually regulated actin filament barbed end-capping and side-binding protein","volume":"285","author":"Dwivedi","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1007\/s12013-009-9047-6","article-title":"The role of vinculin in the regulation of the mechanical properties of cells","volume":"53","author":"Mierke","year":"2009","journal-title":"Cell Biochem. Biophys."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1006\/excr.1996.3451","article-title":"Vinculin promotes cell spreading by mechanically coupling integrins to the cytoskeleton","volume":"231","author":"Ezzell","year":"1997","journal-title":"Exp. Cell Res."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"e106123","DOI":"10.15252\/embj.2020106123","article-title":"Cell softness regulates tumorigenicity and stemness of cancer cells","volume":"40","author":"Lv","year":"2020","journal-title":"EMBO J."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"9208795","DOI":"10.1155\/2018\/9208795","article-title":"Observations of the Effects of Angiotensin II Receptor Blocker on Angiotensin II-Induced Morphological and Mechanical Changes in Renal Tubular Epithelial Cells Using Atomic Force Microscopy","volume":"2018","author":"Kim","year":"2018","journal-title":"BioMed. Res. Int."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1007\/s12668-016-0191-3","article-title":"Discrimination Between Normal and Cancerous Cells Using AFM","volume":"6","author":"Lekka","year":"2016","journal-title":"BioNanoScience"}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/14\/20\/5053\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:54:46Z","timestamp":1760144086000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/14\/20\/5053"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,15]]},"references-count":46,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2022,10]]}},"alternative-id":["cancers14205053"],"URL":"https:\/\/doi.org\/10.3390\/cancers14205053","relation":{},"ISSN":["2072-6694"],"issn-type":[{"value":"2072-6694","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,10,15]]}}}