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EGR-1 forms a complex with YAP-1 and upregulates Bax expression in irradiated prostate carcinoma cells

Oncogene volume 28, pages 1121–1131 (2009)Cite this article

Abstract

In this study, we investigated the functional role of early growth response-1 (Egr1 gene) in the regulation of radiation-induced clonogenic inhibition and apoptosis in p53 wild-type and mutant prostate cancer cells 22Rv1 and DU145, respectively. 22Rv1 cells were more sensitive to irradiation compared with DU145 cells, and the sensitivity was enhanced by overexpression of EGR-1 in both cells. Dominant-negative EGR-1 mutant (dnEGR-1) or repressor of EGR-1, NGFIA binding protein 1 (NAB1), increased radioresistance of these cells. Significant activation of caspases 3 and 9 and Bcl2-associated X (Bax) with increased poly(ADP-ribose) polymerase (PARP) cleavage and cytochrome c release was observed in radiation-exposed EGR-1 overexpressing cells. Gel shift analysis and chloramphenicol acetyl transferase (CAT) reporter assays indicate that EGR-1 transactivates the promoter of the Bax gene. Interaction of EGR-1 and Yes kinase-associated protein 1 (YAP-1) through the WW domain of YAP-1 enhances the transcriptional activity of EGR-1 on the Bax promoter as shown by chromatin immunoprecipitation and reporter assays. Irradiation of PC3 cell xenografts that were treated with adenoviral EGR-1 showed significant regression in tumor volume. These findings establish the radiation-induced pro-apoptotic action of EGR-1, in a p53-independent manner, by directly transactivating Bax, and prove that alters the B-cell CLL/lymphoma 2 (Bcl-2)/Bax ratio as one of the mechanisms resulting in significant activation of caspases, leading to cell death through the novel interaction of EGR-1 with YAP-1.

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Acknowledgements

This work was supported by grants, NIH (RO1 CA78471), PCRP-DOD (PC 970526) and Charlotte Geyer foundation to MMA; post-doctoral training grant, PCRP-DOD (WB1XWH-04-1-0039) to MZ; NIH (DK62345) and Pennsylvania Department of Health Grant (SAP4100037378) to MMS and Welch Foundation Grant (G-1495) to MTB.

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Author notes

  1. M M Shareef, S Gupta & M M Ahmed

    Present address: 8Current address: Department of Radiation Oncology, Sylvester Comprehensive Cancer Center, University of Miami, FL 33136, USA,

  2. M Zagurovskaya, M M Shareef and A Das: These authors contributed equally to this work.

Authors and Affiliations

  1. Weis Center for Research, Geisinger Clinic, Danville, PA, USA

    M Zagurovskaya, M M Shareef, A Reeves, S Gupta, M Sudol & M M Ahmed

  2. Department of Internal Medicine, Virginia Commonwealth University, Richmond, VA, USA

    A Das

  3. Department of Carcinogenesis, M.D. Anderson Cancer Center, University of Texas, Smithville, TX, USA

    M T Bedford

  4. Department of Pathology, Geisinger Clinic, Danville, PA, USA

    J Prichard

  5. Geisinger-Fox Chase Cancer Center, Geisinger Clinic, Wilkes-Barre, PA, USA

    M Mohiuddin

  6. Department of Radiation Oncology, Fox Chase Cancer Center, Philadelphia, PA, USA

    M M Ahmed

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  1. M Zagurovskaya
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Correspondence to M M Ahmed.

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

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Zagurovskaya, M., Shareef, M., Das, A. et al. EGR-1 forms a complex with YAP-1 and upregulates Bax expression in irradiated prostate carcinoma cells. Oncogene 28, 1121–1131 (2009). https://doi.org/10.1038/onc.2008.461

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