GAS5 long non-coding RNA in malignant pleural mesothelioma.

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State: Public
Version: Final published version
Serval ID
serval:BIB_F8AE9A372D8D
Type
Article: article from journal or magazin.
Collection
Publications
Title
GAS5 long non-coding RNA in malignant pleural mesothelioma.
Journal
Molecular Cancer
Author(s)
Renganathan A., Kresoja-Rakic J., Echeverry N., Ziltener G., Vrugt B., Opitz I., Stahel R.A., Felley-Bosco E.
ISSN
1476-4598 (Electronic)
ISSN-L
1476-4598
Publication state
Published
Issued date
2014
Peer-reviewed
Oui
Volume
13
Number
1
Pages
119
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Abstract
BACKGROUND: Malignant pleural mesothelioma (MPM) is an aggressive cancer with short overall survival. Long non-coding RNAs (lncRNA) are a class of RNAs more than 200 nucleotides long that do not code for protein and are part of the 90% of the human genome that is transcribed. Earlier experimental studies in mice showed GAS5 (growth arrest specific transcript 5) gene deletion in asbestos driven mesothelioma. GAS5 encodes for a lncRNA whose function is not well known, but it has been shown to act as glucocorticoid receptor decoy and microRNA "sponge". Our aim was to investigate the possible role of the GAS5 in the growth of MPM.
METHODS: Primary MPM cultures grown in serum-free condition in 3% oxygen or MPM cell lines grown in serum-containing medium were used to investigate the modulation of GAS5 by growth arrest after inhibition of Hedgehog or PI3K/mTOR signalling. Cell cycle length was determined by EdU incorporation assay in doxycycline inducible short hairpinGAS5 clones generated from ZL55SPT cells. Gene expression was quantified by quantitative PCR. To investigate the GAS5 promoter, a 0.77 kb sequence was inserted into a pGL3 reporter vector and luciferase activity was determined after transfection into MPM cells. Localization of GAS5 lncRNA was identified by in situ hybridization. To characterize cells expressing GAS5, expression of podoplanin and Ki-67 was assessed by immunohistochemistry.
RESULTS: GAS5 expression was lower in MPM cell lines compared to normal mesothelial cells. GAS5 was upregulated upon growth arrest induced by inhibition of Hedgehog and PI3K/mTOR signalling in in vitro MPM models. The increase in GAS5 lncRNA was accompanied by increased promoter activity. Silencing of GAS5 increased the expression of glucocorticoid responsive genes glucocorticoid inducible leucine-zipper and serum/glucocorticoid-regulated kinase-1 and shortened the length of the cell cycle. Drug induced growth arrest was associated with GAS5 accumulation in the nuclei. GAS5 was abundant in tumoral quiescent cells and it was correlated to podoplanin expression.
CONCLUSIONS: The observations that GAS5 levels modify cell proliferation in vitro, and that GAS5 expression in MPM tissue is associated with cell quiescence and podoplanin expression support a role of GAS5 in MPM biology.
Keywords
Cell Cycle/genetics, Cell Line, Tumor, Cell Proliferation, Gene Expression Regulation, Neoplastic, Genes, Reporter, Hedgehog Proteins/genetics, Hedgehog Proteins/metabolism, Humans, Ki-67 Antigen/genetics, Ki-67 Antigen/metabolism, Luciferases/genetics, Luciferases/metabolism, Lung Neoplasms/genetics, Lung Neoplasms/metabolism, Membrane Glycoproteins/genetics, Membrane Glycoproteins/metabolism, Mesothelioma/genetics, Mesothelioma/metabolism, Phosphatidylinositol 3-Kinases/antagonists & inhibitors, Phosphatidylinositol 3-Kinases/genetics, Primary Cell Culture, Promoter Regions, Genetic, Protein Kinase Inhibitors/pharmacology, RNA, Long Noncoding/antagonists & inhibitors, RNA, Long Noncoding/genetics, RNA, Small Interfering/genetics, RNA, Small Interfering/metabolism, Signal Transduction, TOR Serine-Threonine Kinases/antagonists & inhibitors, TOR Serine-Threonine Kinases/genetics
Pubmed
Web of science
Open Access
Yes
Create date
03/08/2014 11:11
Last modification date
20/08/2019 16:24
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