Markus Wehland

Alterations of Growth and Focal Adhesion Molecules in Human Breast Cancer Cells Exposed to the Random Positioning Machine

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

  • Jayashree Sahana
  • Thomas J Corydon
  • Markus Wehland
  • Marcus Krüger, Otto von Guericke University Magdeburg
  • ,
  • Sascha Kopp, Otto von Guericke University Magdeburg
  • ,
  • Daniela Melnik, Otto von Guericke University Magdeburg
  • ,
  • Stefan Kahlert, Otto von Guericke University Magdeburg
  • ,
  • Borna Relja, Otto von Guericke University Magdeburg
  • ,
  • Manfred Infanger, Otto von Guericke University Magdeburg
  • ,
  • Daniela Grimm

In this study, we evaluated changes in focal adhesions (FAs) in two types of breast cancer cell (BCC) lines (differentiated MCF-7 and the triple-negative MDA-MB-231 cell line) exposed to simulated microgravity (s-μg) created by a random positioning machine (RPM) for 24 h. After exposure, the BCC changed their growth behavior and exhibited two phenotypes in RPM samples: one portion of the cells grew as a normal two-dimensional monolayer [adherent (AD) BCC], while the other portion formed three-dimensional (3D) multicellular spheroids (MCS). After 1 h and 30 min (MDA-MB-231) and 1 h 40 min (MCF-7), the MCS adhered completely to the slide flask bottom. After 2 h, MDA-MB-231 MCS cells started to migrate, and after 6 h, a large number of the cells had left the MCS and continued to grow in a scattered pattern, whereas MCF-7 cells were growing as a confluent monolayer after 6 h and 24 h. We investigated the genes associated with the cytoskeleton, the extracellular matrix and FAs. ACTB, TUBB, FN1, FAK1, and PXN gene expression patterns were not significantly changed in MDA-MB-231 cells, but we observed a down-regulation of LAMA3, ITGB1 mRNAs in AD cells and of ITGB1, TLN1 and VCL mRNAs in MDA-MB-231 MCS. RPM-exposed MCF-7 cells revealed a down-regulation in the gene expression of FAK1, PXN, TLN1, VCL and CDH1 in AD cells and PXN, TLN and CDH1 in MCS. An interaction analysis of the examined genes involved in 3D growth and adhesion indicated a central role of fibronectin, vinculin, and E-cadherin. Live cell imaging of eGFP-vinculin in MCF-7 cells confirmed these findings. β-catenin-transfected MCF-7 cells revealed a nuclear expression in 1g and RPM-AD cells. The target genes BCL9, MYC and JUN of the Wnt/β-catenin signaling pathway were differentially expressed in RPM-exposed MCF-7 cells. These findings suggest that vinculin and β-catenin are key mediators of BCC to form MCS during 24 h of RPM-exposure.

Original languageEnglish
Article number672098
JournalFrontiers in Cell and Developmental Biology
Volume9
Number of pages17
ISSN2296-634X
DOIs
Publication statusPublished - 2021

Bibliographical note

Copyright © 2021 Sahana, Corydon, Wehland, Krüger, Kopp, Melnik, Kahlert, Relja, Infanger and Grimm.

See relations at Aarhus University Citationformats

ID: 219990730