Molecular Insights into Division of Single Human Cancer Cells in On-Chip Transparent Microtubes.
In vivo, mammalian cells proliferate within 3D environments consisting of numerous microcavities and channels, which contain a variety of chemical and physical cues. External environments often differ between normal and pathological states, such as the unique spatial constraints that metastasizing cancer cells experience as they circulate the vasculature through arterioles and narrow capillaries, where they can divide and acquire elongated cylindrical shapes. While metastatic tumors cause most cancer deaths, factors impacting early cancer cell proliferation inside the vasculature and those that can promote the formation of secondary tumors remain largely unknown. Prior studies investigating confined mitosis have mainly used 2D cell culture systems. Here, we mimic aspects of metastasizing tumor cells dividing inside blood capillaries by investigating single-cell divisions of living human cancer cells, trapped inside 3D rolled-up, transparent nanomembranes. We assess the molecular effects of tubular confinement on key mitotic features, using optical high- and super-resolution microscopy. Our experiments show that tubular confinement affects the morphology and dynamics of the mitotic spindle, chromosome arrangements, and the organization of the cell cortex. Moreover, we reveal that membrane blebbing and/or associated processes act as a potential genome-safety mechanism, limiting the extent of genomic instability caused by mitosis in confined circumstances, especially in tubular 3D microenvironments. Collectively, our study demonstrates the potential of rolled-up nanomembranes for gaining molecular insights into key cellular events occurring in tubular 3D microenvironments in vivo.
Institute for Integrative Nanosciences, IFW Dresden , Helmholtzstr. 20, D-01069 Dresden, Germany.
Wang
W
Xi
The Gurdon Institute and Departments of Biochemistry, Genetics and Pharmacology, University of Cambridge , Tennis Court Road, Cambridge CB2 1QN, United Kingdom.
Christine K
CK
Schmidt
Institute for Integrative Nanosciences, IFW Dresden , Helmholtzstr. 20, D-01069 Dresden, Germany.
Samuel
S
Sanchez
Department of Chemical and Biomolecular Engineering, Johns Hopkins University , Baltimore, Maryland 21218, United States.
David H
DH
Gracias
The Gurdon Institute and Departments of Biochemistry, Genetics and Pharmacology, University of Cambridge , Tennis Court Road, Cambridge CB2 1QN, United Kingdom.
Rafael E
RE
Carazo-Salas
The Gurdon Institute and Departments of Biochemistry, Genetics and Pharmacology, University of Cambridge , Tennis Court Road, Cambridge CB2 1QN, United Kingdom.
Richard
R
Butler
The Gurdon Institute and Departments of Biochemistry, Genetics and Pharmacology, University of Cambridge , Tennis Court Road, Cambridge CB2 1QN, United Kingdom.
Nicola
N
Lawrence
The Gurdon Institute and Departments of Biochemistry, Genetics and Pharmacology, University of Cambridge , Tennis Court Road, Cambridge CB2 1QN, United Kingdom.|The Wellcome Trust Sanger Institute , Hinxton, Cambridge CB10 1SA, United Kingdom.
Stephen P
SP
Jackson
Institute for Integrative Nanosciences, IFW Dresden , Helmholtzstr. 20, D-01069 Dresden, Germany.|Material Systems for Nanoelectronics, Chemnitz University of Technology , Reichenhainer Str. 70, D-09107 Chemnitz, Germany.|Center for Advancing Electronics Dresden, Dresden University of Technology , Georg-Schumann-Str. 11, 01187 Dresden, Germany.
Oliver G
OG
Schmidt
D000818:Animals; D049109:Cell Proliferation; D006801:Humans; D008853:Microscopy; D008938:Mitosis; D049329:Nanostructures; D043942:Nanotubes; D009369:Neoplasms
3D cell culture; actin cortex; chromosome segregation; membrane blebbing; metastasis; mitosis; rolled-up nanofilms/membranes