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  • Original Paper
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Evidence that Sp1 positively and Sp3 negatively regulate and androgen does not directly regulate functional tumor suppressor 15-lipoxygenase 2 (15-LOX2) gene expression in normal human prostate epithelial cells

Oncogene volume 23, pages 6942–6953 (2004)Cite this article

Abstract

In this project, we studied the gene regulation of 15-lipoxygenase 2 (15-LOX2), the most abundant arachidonate-metabolizing LOX in adult human prostate and a negative cell-cycle regulator in normal human prostate (NHP) epithelial cells. Through detailed in silico promoter examination and promoter deletion and activity analysis, we found that several Sp1 sites (i.e., three GC boxes and one CACCC box) in the proximal promoter region play a critical role in regulating 15-LOX2 expression in NHP cells. Several pieces of evidence further suggest that the Sp1 and Sp3 proteins play a physiologically important role in positively and negatively regulating the 15-LOX2 gene expression, respectively. First, mutations in the GC boxes affected the 15-LOX2 promoter activity. Second, both Sp1 and Sp3 proteins were detected in the protein complexes that bound the GC boxes revealed by electrophoretic mobility shift assay. Third, importantly, inhibition of Sp1 activity or overexpression of Sp3 both inhibited the endogenous 15-LOX2 mRNA expression. Since 15-LOX2 is normally expressed in the prostate luminal epithelial cells, we subsequently explored whether androgen/androgen receptor may directly regulate its gene expression. The results indicate that androgen does not directly regulate 15-LOX2 gene expression. Together, these observations provide insight on how 15-LOX2 gene expression may be regulated in NHP cells.

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Abbreviations

AA:

arachidonic acid

AR:

androgen receptor

DN:

dominant negative

EMSA:

electrophoretic mobility shift assay

LOX:

lipoxygenase

15-LOX2:

15-lipoxygenase 2

15-LOX2sv-a/b/c:

15-LOX2 splice variant a, b or c

MMA:

mithramycin A

NE:

nuclear extract

NHP:

normal human prostate epithelial cells

PCa:

prostate cancer

RLU:

relative luciferase unit

TSS:

transcription start site

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Acknowledgements

We thank D Chopra for NHP2 cells, G Thiel for DN-Sp1, L W-K Chung for AR, R Matusik for the ARR2PB promoter, J-J Shen and Molecular Biology Core for assistance in sequencing, and members of the Tang lab for helpful discussions. This work is supported, in part, by R01 CA90297, ACS Grant RSG-03-163-01-MGO, DOD grant DAMD17-03-1-0137, and NIEHS Center Grant ES07784. SL Friedman is supported by R01 DK 37340. CJ Maldonado is supported by NIH post-doctoral training grant T32 CA09480-16. D Chandra is supported by DOD Postdoctoral Traineeship Award DAMD17-02-1-0083. B Bhatia is a graduate student in GSBS program.

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  1. Carlos J Maldonado

    Present address: Biotechnology Transition Cell Molecular Epidemiology Branch, AFIOH/SDEM, 2730 Louis Bauer Dr., Brooks AFB, San Antonio, TX, 78235, USA

Authors and Affiliations

  1. Department of Carcinogenesis, Science Park-Research Division, The University of Texas MD Anderson Cancer Center, 1808 Park Rd. 1C, Smithville, 78957, TX, USA

    Shaohua Tang, Bobby Bhatia, Jianjun Zhou, Carlos J Maldonado, Dhyan Chandra, Eunjung Kim, Susan M Fischer, Andrew P Butler & Dean G Tang

  2. Department of Medicine, Division of Liver Diseases, Mount Sinai School of Medicine, 1425 Madison Ave., New York, 10029, NY, USA

    Scott L Friedman

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  1. Shaohua Tang
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Correspondence to Dean G Tang.

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Tang, S., Bhatia, B., Zhou, J. et al. Evidence that Sp1 positively and Sp3 negatively regulate and androgen does not directly regulate functional tumor suppressor 15-lipoxygenase 2 (15-LOX2) gene expression in normal human prostate epithelial cells. Oncogene 23, 6942–6953 (2004). https://doi.org/10.1038/sj.onc.1207913

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