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Integrin β1, osmosensing, and chemoresistance in mouse ehrlich carcinoma cells

Sørensen, Belinda Halling; Rasmussen, Line Jee Hartmann; Broberg, Bjørn Sindballe; Klausen, Thomas Kjær; Sauter, Daniel Peter Rafael; Lambert, Ian Henry; Aspberg, Anders LU and Hoffmann, Else Kay (2015) In Cellular Physiology and Biochemistry 36(1). p.111-132
Abstract

Background/Aims: Altered expression of the integrin family of cell adhesion receptors has been associated with initiation, progression, and metastasis of solid tumors as well as in the development of chemoresistance. Here, we investigated the role of integrins, in particular integrin β1, in cell volume regulation and drug-induced apoptosis in adherent and non-adherent Ehrlich ascites cell lines. Methods: Adhesion phenotypes were verified by colorimetric cell-adhesion-assay. Quantitative real-time PCR and western blot were used to compare expression levels of integrin subunits. Small interfering RNA was used to silence integrin β1 expression. Regulatory volume decrease (RVD) after cell swelling was studied with... (More)

Background/Aims: Altered expression of the integrin family of cell adhesion receptors has been associated with initiation, progression, and metastasis of solid tumors as well as in the development of chemoresistance. Here, we investigated the role of integrins, in particular integrin β1, in cell volume regulation and drug-induced apoptosis in adherent and non-adherent Ehrlich ascites cell lines. Methods: Adhesion phenotypes were verified by colorimetric cell-adhesion-assay. Quantitative real-time PCR and western blot were used to compare expression levels of integrin subunits. Small interfering RNA was used to silence integrin β1 expression. Regulatory volume decrease (RVD) after cell swelling was studied with calcein-fluorescence-self-quenching and Coulter counter analysis. Taurine efflux was estimated with tracer technique. Caspase assay was used to determine apoptosis. Results: We show that adherent cells have stronger fibronectin binding and a significantly increased expression of integrin α5, αv, and β1 at mRNA and protein level, compared to non-adherent cells. Knockdown of integrin β1 reduced RVD of the adherent but not of the non-adherent cells. Efflux of taurine was unaffected. In contrast to non-adherent, adherent cells exhibited chemoresistance to chemotherapeutic drugs (cisplatin and gemcitabine). However, knockdown of integrin β1 promoted cisplatin-induced caspase activity in adherent cells. Conclusion: Our data identifies integrin β1 as a part of the osmosensing machinery and regulator of cisplatin resistance in adherent Ehrlich cells.

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author
publishing date
type
Contribution to journal
publication status
published
keywords
Cisplatin, Ehrlich Ascites Tumor Cells, Gemcitabine, Hypotonic cell swelling, Integrin, Multi-drug-resistance
in
Cellular Physiology and Biochemistry
volume
36
issue
1
pages
111 - 132
publisher
Karger
external identifiers
  • scopus:84928752956
ISSN
1015-8987
DOI
10.1159/000374057
language
English
LU publication?
no
id
3633340c-dee2-4f9b-bc59-483151cf51f9
date added to LUP
2016-12-06 10:05:18
date last changed
2017-11-05 05:10:36
@article{3633340c-dee2-4f9b-bc59-483151cf51f9,
  abstract     = {<p>Background/Aims: Altered expression of the integrin family of cell adhesion receptors has been associated with initiation, progression, and metastasis of solid tumors as well as in the development of chemoresistance. Here, we investigated the role of integrins, in particular integrin β<sub>1</sub>, in cell volume regulation and drug-induced apoptosis in adherent and non-adherent Ehrlich ascites cell lines. Methods: Adhesion phenotypes were verified by colorimetric cell-adhesion-assay. Quantitative real-time PCR and western blot were used to compare expression levels of integrin subunits. Small interfering RNA was used to silence integrin β<sub>1</sub> expression. Regulatory volume decrease (RVD) after cell swelling was studied with calcein-fluorescence-self-quenching and Coulter counter analysis. Taurine efflux was estimated with tracer technique. Caspase assay was used to determine apoptosis. Results: We show that adherent cells have stronger fibronectin binding and a significantly increased expression of integrin α<sub>5</sub>, α<sub>v</sub>, and β<sub>1</sub> at mRNA and protein level, compared to non-adherent cells. Knockdown of integrin β<sub>1</sub> reduced RVD of the adherent but not of the non-adherent cells. Efflux of taurine was unaffected. In contrast to non-adherent, adherent cells exhibited chemoresistance to chemotherapeutic drugs (cisplatin and gemcitabine). However, knockdown of integrin β<sub>1</sub> promoted cisplatin-induced caspase activity in adherent cells. Conclusion: Our data identifies integrin β<sub>1</sub> as a part of the osmosensing machinery and regulator of cisplatin resistance in adherent Ehrlich cells.</p>},
  author       = {Sørensen, Belinda Halling and Rasmussen, Line Jee Hartmann and Broberg, Bjørn Sindballe and Klausen, Thomas Kjær and Sauter, Daniel Peter Rafael and Lambert, Ian Henry and Aspberg, Anders and Hoffmann, Else Kay},
  issn         = {1015-8987},
  keyword      = {Cisplatin,Ehrlich Ascites Tumor Cells,Gemcitabine,Hypotonic cell swelling,Integrin,Multi-drug-resistance},
  language     = {eng},
  month        = {05},
  number       = {1},
  pages        = {111--132},
  publisher    = {Karger},
  series       = {Cellular Physiology and Biochemistry},
  title        = {Integrin β<sub>1</sub>, osmosensing, and chemoresistance in mouse ehrlich carcinoma cells},
  url          = {http://dx.doi.org/10.1159/000374057},
  volume       = {36},
  year         = {2015},
}