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

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

Abstract 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 determin...

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Autores principales: Sascha Kopp, Jayashree Sahana, Tawhidul Islam, Asbjørn Graver Petersen, Johann Bauer, Thomas J. Corydon, Herbert Schulz, Kathrin Saar, Norbert Huebner, Lasse Slumstrup, Stefan Riwaldt, Markus Wehland, Manfred Infanger, Ronald Luetzenberg, Daniela Grimm
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Publicado: Nature Portfolio 2018
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Acceso en línea:https://doaj.org/article/4792f639a36f4440a59f0c9fd262c315
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spelling oai:doaj.org-article:4792f639a36f4440a59f0c9fd262c3152021-12-02T15:08:29ZThe role of NFκB in spheroid formation of human breast cancer cells cultured on the Random Positioning Machine10.1038/s41598-017-18556-82045-2322https://doaj.org/article/4792f639a36f4440a59f0c9fd262c3152018-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-18556-8https://doaj.org/toc/2045-2322Abstract 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.Sascha KoppJayashree SahanaTawhidul IslamAsbjørn Graver PetersenJohann BauerThomas J. CorydonHerbert SchulzKathrin SaarNorbert HuebnerLasse SlumstrupStefan RiwaldtMarkus WehlandManfred InfangerRonald LuetzenbergDaniela GrimmNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-17 (2018)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Sascha Kopp
Jayashree Sahana
Tawhidul Islam
Asbjørn Graver Petersen
Johann Bauer
Thomas J. Corydon
Herbert Schulz
Kathrin Saar
Norbert Huebner
Lasse Slumstrup
Stefan Riwaldt
Markus Wehland
Manfred Infanger
Ronald Luetzenberg
Daniela Grimm
The role of NFκB in spheroid formation of human breast cancer cells cultured on the Random Positioning Machine
description Abstract 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.
format article
author Sascha Kopp
Jayashree Sahana
Tawhidul Islam
Asbjørn Graver Petersen
Johann Bauer
Thomas J. Corydon
Herbert Schulz
Kathrin Saar
Norbert Huebner
Lasse Slumstrup
Stefan Riwaldt
Markus Wehland
Manfred Infanger
Ronald Luetzenberg
Daniela Grimm
author_facet Sascha Kopp
Jayashree Sahana
Tawhidul Islam
Asbjørn Graver Petersen
Johann Bauer
Thomas J. Corydon
Herbert Schulz
Kathrin Saar
Norbert Huebner
Lasse Slumstrup
Stefan Riwaldt
Markus Wehland
Manfred Infanger
Ronald Luetzenberg
Daniela Grimm
author_sort Sascha Kopp
title The role of NFκB in spheroid formation of human breast cancer cells cultured on the Random Positioning Machine
title_short The role of NFκB in spheroid formation of human breast cancer cells cultured on the Random Positioning Machine
title_full The role of NFκB in spheroid formation of human breast cancer cells cultured on the Random Positioning Machine
title_fullStr The role of NFκB in spheroid formation of human breast cancer cells cultured on the Random Positioning Machine
title_full_unstemmed The role of NFκB in spheroid formation of human breast cancer cells cultured on the Random Positioning Machine
title_sort role of nfκb in spheroid formation of human breast cancer cells cultured on the random positioning machine
publisher Nature Portfolio
publishDate 2018
url https://doaj.org/article/4792f639a36f4440a59f0c9fd262c315
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