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PMID: 11687610 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Design of a retroviral-mediated ecdysone-inducible system and its application to the expression profiling of the PTEN tumor suppressor.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 98 ·No. 23 ·2001-11-06 ·Pages 13043-8

Stolarov J, Chang K, Reiner A, Rodgers L, Hannon GJ, Wigler MH, Mittal V

Abstract

We have engineered the ecdysone-inducible mammalian expression system for general retroviral delivery to cultured mammalian cells. We inducibly expressed PTEN in the glioblastoma cell line, U87MG, lacking this gene. Because nearly all cells are recruited on induction, we find both up- and down-regulated genes by cDNA microarray analysis. The changes we see are similar to those observed after treatment with LY294002, an inhibitor of phosphatidylinositol 3-OH kinase, fully consistent with the model that PTEN antagonizes phosphatidylinositol 3-OH kinase. Both treatments result in suppressed expression of the transforming growth factor (TGF)-beta gene and the genes of the cholesterol biosynthesis pathway. Our results illustrate the power of using a fully inducible expression system in conjunction with cDNA microarray analysis for exploring gene function.

MeSH Terms

Ecdysone/biosynthesis Flow Cytometry Gene Expression Profiling Genes, Tumor Suppressor Genetic Vectors Humans PTEN Phosphohydrolase Phosphoric Monoester Hydrolases/genetics Retroviridae/genetics Transcription, Genetic Tumor Cells, Cultured Tumor Suppressor Proteins/genetics

Chemicals

Tumor Suppressor Proteins Ecdysone Phosphoric Monoester Hydrolases PTEN Phosphohydrolase PTEN protein, human

Authors & Affiliations

7 authors, click to expand affiliations / ORCID

Stolarov J

Cold Spring Harbor Laboratory, 1 Bungtown Road, P.O. Box 100, Cold Spring Harbor, NY 11724, USA.

Chang K

Reiner A

Rodgers L

Hannon G J

Wigler M H

Mittal V

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Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.221450598

Published

2001-11-06

Epub

2001-00-30

Pages

13043-8

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC60821

Subset

Grants

NCI NIH HHS · R01 CA078544 · United States

NCI NIH HHS · 5R01-CA78544 · United States

External Links

PubMed source DOI: 10.1073/pnas.221450598 PMC Full Text View journal details

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Design of a retroviral-mediated ecdysone-inducible system and its application to the expression profiling of the PTEN tumor suppressor.

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