Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Mechanisms of Basement Membrane Micro-Perforation During Cancer Cell Invasion Into a 3D Collagen Gel

Version 1 : Received: 10 August 2022 / Approved: 12 August 2022 / Online: 12 August 2022 (12:55:21 CEST)

A peer-reviewed article of this Preprint also exists.

Nazari, S.S.; Doyle, A.D.; Yamada, K.M. Mechanisms of Basement Membrane Micro-Perforation during Cancer Cell Invasion into a 3D Collagen Gel. Gels 2022, 8, 567. Nazari, S.S.; Doyle, A.D.; Yamada, K.M. Mechanisms of Basement Membrane Micro-Perforation during Cancer Cell Invasion into a 3D Collagen Gel. Gels 2022, 8, 567.

Journal reference: Gels 2022, 8, 567
DOI: 10.3390/gels8090567

Abstract

Cancer invasion through basement membranes represents the initial step of tumor dissemination and metastasis. However, little is known about how human cancer cells breach basement membranes. Here, we used a 3-dimensional in vitro invasion model consisting of cancer spheroids encapsulated by a basement membrane and embedded in 3D collagen gels to visualize the early events of cancer invasion by confocal microscopy and live-cell imaging. Human breast cancer cells generated large numbers of basement membrane perforations, or holes, of varying sizes that expanded over time during cell invasion. We used a wide variety of small molecule inhibitors to probe the mechanisms of basement membrane perforation and hole expansion. Protease inhibitor treatment (BB94), led to a 63% decrease in perforation size. After myosin II inhibition (blebbistatin), basement membrane perforation area decreased by only 15%. These treatments produced correspondingly decreased cellular breaching events. Interestingly, inhibition of actin polymerization dramatically decreased basement membrane perforation by 80% and blocked invasion. Our findings suggest that human cancer cells can primarily use proteolysis and actin polymerization to perforate the BM and to expand perforations for basement membrane breaching, with a relatively small contribution from myosin II contractility.

Keywords

hydrogel; 3D-culture; Imaging; Cell-matrix; proteases; matrix metalloproteinases; actin polymerization; contractility

Subject

LIFE SCIENCES, Cell & Developmental Biology

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