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Unphosphorylated STAT6 contributes to constitutive cyclooxygenase-2 expression in human non-small cell lung cancer

Oncogene volume 26, pages 4253–4260 (2007)Cite this article

Abstract

Cyclooxygenase-2 (COX-2) is frequently overexpressed in human cancers and contributes to the malignant phenotype. Our data indicate unphosphorylated signal transducers and activators of transcription 6 (STAT6) may transcriptionally upregulate COX-2 expression and protect against apoptosis in NSCLC cells. In A427 and H2122, NSCLC cell lines that constitutively express COX-2, only unphosphorylated STAT6 was detectable by western blot, thus, all of the following STAT6-dependent effects are attributed to the unphosphorylated protein. In both cell lines, small-interfering RNA-mediated knockdown of STAT6 or stable expression of dominant-negative STAT6 decreased COX-2 expression. In contrast, transfection with a phosphorylation-deficient mutant STAT6 increased COX-2 levels. Immunofluorescent staining revealed the presence of STAT6 in H2122 nuclei, suggesting a direct role in gene regulation for the unphosphorylated protein. Consistent with this hypothesis, unphosphorylated STAT6 increased luciferase expression from a COX-2 promoter reporter construct. STAT6 co-immunoprecipitated with the transcriptional co-activator, p300, and chromatin immunoprecipitation assays demonstrated that these proteins bind a consensus STAT6 binding site located within the COX-2 promoter. STAT6 DNA-binding specificity was confirmed by electrophoretic mobility shift assay. As COX-2 over-expression has been clearly linked to apoptosis resistance and other hallmarks of malignancy, these findings suggest a novel role of unphosphorylated STAT6 in the pathogenesis of non-small cell lung cancer.

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Acknowledgements

We thank Dr Katherine Peebles for reviewing the manuscript. This work was supported by University of California at Los Angeles Specialized Program of Research Excellence In Lung Cancer NIH grant P50 CA90388, UC Tobacco-Related Disease Research Program 13RT-0031, NIH RO1 CA111851 and Merit Review Research Funds from the Department of Veterans Affairs.

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Authors and Affiliations

  1. Division of Pulmonary and Critical Care Medicine, Department of Medicine, Lung Cancer Research Program of the Jonsson Comprehensive Cancer Center, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA, USA

    X Cui, L Zhang, J Luo, S Hazra, N Cacalano & S M Dubinett

  2. Department of Pathology, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA, USA

    A Rajasekaran & S M Dubinett

  3. Department of Radiation Oncology, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA, USA

    N Cacalano

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  1. X Cui
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Correspondence to S M Dubinett.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Cui, X., Zhang, L., Luo, J. et al. Unphosphorylated STAT6 contributes to constitutive cyclooxygenase-2 expression in human non-small cell lung cancer. Oncogene 26, 4253–4260 (2007). https://doi.org/10.1038/sj.onc.1210222

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