During metastasis, cancer cells navigate through harsh conditions, including various mechanical forces in the bloodstream, highlighting the need to understand the impact of mechanical and shear stresses on cancer cells. To overcome the current methodological limitations of such research, here we present a new device that replicates similar conditions by applying shear stress on cultured cells. The device provides a less complex, easily accessible alternative to traditional fluidics while generating fluid shear stress values comparable to those in human veins and capillaries. The device allows analyses of large cell numbers in standard cell culture flasks and incubators. Using this device to explore the shear stress-induced responses of various human cell lines, we discovered a previously unknown, reversible pre-cytokinetic block occurring in cells that lose anchorage during mitosis and are kept under constant shear stress. Notably, some cancer cell lines appear to bypass this unorthodox cell-cycle block, suggesting its role as a safety checkpoint to restrict the proliferation of cancer cells in the bloodstream and their overall spreading potential. These findings provide new insights into the diverse responses of normal and cancer cells to shear stress and highlight the potential of our technology for research on circulating tumor cells and metastatic spread.

Danish Cancer Institute Danish Cancer Society Copenhagen Denmark

Department of Biochemistry Faculty of Medicine Masaryk University Kamenice 5 Brno 625 00 Czech Republic

Department of Biophysics of Immune System Institute of Biophysics of the Czech Academy of Sciences Kralovopolska 135 Brno Czech Republic

Department of Optics Faculty of Science Palacky University Olomouc Czech Republic

Division of Genome Biology Department of Medical Biochemistry and Biophysics Science for Life Laboratory Karolinska Institute Stockholm Sweden

Faculty of Mechanical Engineering Jan Evangelista Purkyně University Ústí Nad Labem Czech Republic

International Clinical Research Center St Anne's University Hospital Brno Brno Czech Republic

Laboratory of Genome Integrity Faculty of Medicine and Dentistry Institute of Molecular and Translational Medicine Palacky University Olomouc Czech Republic

Zobrazit více v PubMed

Chaffer, C. L. & Weinberg, R. A. A perspective on cancer cell metastasis. Science331, 1559–1564 (2011). PubMed

Huang, Q. et al. Fluid shear stress and tumor metastasis. Am. J. Cancer Res.8, 763–777 (2018). PubMed PMC

Follain, G. et al. Fluids and their mechanics in tumour transit: Shaping metastasis. Nat. Rev. Cancer20, 107–124 (2020). PubMed

Fan, R. et al. Circulatory shear flow alters the viability and proliferation of circulating colon cancer cells. Sci. Rep.6, 27073 (2016). PubMed PMC

Sinkala, E. et al. Profiling protein expression in circulating tumour cells using microfluidic western blotting. Nat. Commun.8, 14622 (2017). PubMed PMC

Abouleila, Y. et al. Live single cell mass spectrometry reveals cancer-specific metabolic profiles of circulating tumor cells. Cancer Sci.110, 697–706 (2019). PubMed PMC

Luo, C.-W., Wu, C.-C. & Ch’ang, H.-J. Radiation sensitization of tumor cells induced by shear stress: The roles of integrins and FAK. Biochim. Biophys. Acta1843, 2129–2137 (2014). PubMed

Chiu, J.-J. & Chien, S. Effects of disturbed flow on vascular endothelium: Pathophysiological basis and clinical perspectives. Physiol. Rev.91, 327–387 (2011). PubMed PMC

Zhou, J., Li, Y. S. & Chien, S. Shear stress-initiated signaling and its regulation of endothelial function. Arterioscler. Thromb. Vasc. Biol.34, 2191 (2014). PubMed PMC

Turitto, V. T. Blood viscosity, mass transport, and thrombogenesis. Prog. Hemost. Thromb.6, 139–177 (1982). PubMed

Shemesh, J. et al. Flow-induced stress on adherent cells in microfluidic devices. Lab. Chip15, 4114–4127 (2015). PubMed

World, C. J., Garin, G. & Berk, B. Vascular shear stress and activation of inflammatory genes. Curr. Atheroscler. Rep.8, 240–244 (2006). PubMed

Tanaka, K., Joshi, D., Timalsina, S. & Schwartz, M. A. Early events in endothelial flow sensing. Cytoskelet. Hoboken NJ78, 217–231 (2021). PubMed

Hahn, C. & Schwartz, M. A. The role of cellular adaptation to mechanical forces in atherosclerosis. Arterioscler. Thromb. Vasc. Biol.28, 2101–2107 (2008). PubMed PMC

Nagel, T., Resnick, N., Atkinson, W. J., Dewey, C. F. & Gimbrone, M. A. Shear stress selectively upregulates intercellular adhesion molecule-1 expression in cultured human vascular endothelial cells. J. Clin. Invest.94, 885–891 (1994). PubMed PMC

Bao, X., Lu, C. & Frangos, J. A. Mechanism of temporal gradients in shear-induced ERK1/2 activation and proliferation in endothelial cells. Am. J. Physiol. Heart Circ. Physiol.281, H22-29 (2001). PubMed

Ishida, T., Peterson, T. E., Kovach, N. L. & Berk, B. C. MAP kinase activation by flow in endothelial cells. Circ. Res.79, 310–316 (1996). PubMed

Mengistu, M., Brotzman, H., Ghadiali, S. & Lowe-Krentz, L. Fluid shear stress-induced JNK activity leads to actin remodeling for cell alignment. J. Cell. Physiol.226, 110–121 (2011). PubMed PMC

Thayse, K., Kindt, N., Laurent, S. & Carlier, S. VCAM-1 target in non-invasive imaging for the detection of atherosclerotic plaques. Biology9, 368 (2020). PubMed PMC

Taubenberger, A. V., Baum, B. & Matthews, H. K. The mechanics of mitotic cell rounding. Front. Cell Dev. Biol.8, 687 (2020). PubMed PMC

Fujiwara, T. et al. Cytokinesis failure generating tetraploids promotes tumorigenesis in p53-null cells. Nature437, 1043–1047 (2005). PubMed

Černík, M. et al. Luminal surface plasma treatment of closed cylindrical microchannels: A tool toward the creation of on-chip vascular endothelium. ACS Biomater. Sci. Eng.9, 2755–2763 (2023). PubMed PMC

Schneider, C. A., Rasband, W. S. & Eliceiri, K. W. NIH Image to ImageJ: 25 years of image analysis. Nat. Methods9, 671–675 (2012). PubMed PMC

Kudlova, N. et al. An efficient, non-invasive approach for in-vivo sampling of hair follicles: Design and applications in monitoring DNA damage and aging. Aging13, 25004–25024 (2021). PubMed PMC