Aarhus Universitet

Jayashree Sahana

The role of NFκB in spheroid formation of human breast cancer cells cultured on the Random Positioning Machine

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DOI

  • Sascha Kopp, Clinic and Policlinic for Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke-University, 39120 Magdeburg, Germany. sascha.kopp@med.ovgu.de.
  • ,
  • Jayashree Sahana
  • Tawhidul Islam
  • ,
  • Asbjørn Graver Petersen
  • Johann Bauer, Max-Planck-Institute for Biochemistry, Scientific Information Services, 82152 Martinsried, Germany. jbauer@biochem.mpg.de.
  • ,
  • Thomas J Corydon
  • Herbert Schulz, Cologne Center for Genomics, University of Cologne, D-50931, Cologne, Germany.
  • ,
  • Kathrin Saar, Max-Delbrück-Center for Molecular Medicine, D-13092, Berlin-Buch, Germany.
  • ,
  • Norbert Huebner, Max-Delbrück-Center for Molecular Medicine, D-13092, Berlin-Buch, Germany.
  • ,
  • Lasse Slumstrup
  • ,
  • Stefan Riwaldt
  • Markus Wehland, Clinic and Policlinic for Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke-University, 39120 Magdeburg, Germany. sascha.kopp@med.ovgu.de.
  • ,
  • Manfred Infanger, Clinic and Policlinic for Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke-University, 39120 Magdeburg, Germany. sascha.kopp@med.ovgu.de.
  • ,
  • Ronald Luetzenberg, Clinic and Policlinic for Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke-University, 39120 Magdeburg, Germany. sascha.kopp@med.ovgu.de.
  • ,
  • Daniela Grimm

Human MCF-7 breast cancer cells were exposed to a Random Positioning Machine (RPM). After 24 hours (h) the cells grew either adherently within a monolayer (AD) or within multicellular spheroids (MCS). AD and MCS populations were separately harvested, their cellular differences were determined performing qPCR on genes, which were differently expressed in AD and MCS cells. Gene array technology was applied to detect RPM-sensitive genes in MCF-7 cells after 24 h. Furthermore, the capability to form multicellular spheroids in vitro was compared with the intracellular distribution of NF-kappaB (NFκB) p65. NFκB was equally distributed in static control cells, but predominantly localized in the cytoplasm in AD cells and nucleus in MCS cells exposed to the RPM. Gene array analyses revealed a more than 2-fold change of only 23 genes including some whose products are affected by oxygen levels or regulate glycolysis. Significant upregulations of the mRNAs of enzymes degrading heme, of ANXA1, ANXA2, CTGF, CAV2 and ICAM1, as well as of FAS, Casp8, BAX, p53, CYC1 and PARP1 were observed in MCS cells as compared with 1g-control and AD cells. An interaction analysis of 47 investigated genes suggested that HMOX-1 and NFκB variants are activated, when multicellular spheroids are formed.

OriginalsprogEngelsk
Artikelnummer921
TidsskriftScientific Reports
Vol/bind8
Nummer1
ISSN2045-2322
DOI
StatusUdgivet - 17 jan. 2018

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