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Originally published In Press as doi:10.1074/jbc.M105947200 on October 29, 2001

J. Biol. Chem., Vol. 277, Issue 4, 2468-2476, January 25, 2002
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Regulation of Human Osteocalcin Promoter in Hormone-independent Human Prostate Cancer Cells*

Fan YeungDagger §, Wai K. LawDagger , Ching-Hua Yeh||, Jennifer J. Westendorf**, Ye ZhangDagger , Ruoxiang WangDagger Dagger , Chinghai Kao§§, and Leland W. K. ChungDagger §Dagger Dagger ¶¶

From the Departments of Dagger  Urology, § Cell Biology,  Biochemistry and Molecular Genetics, and || Orthopedic Surgery, University of Virginia, Charlottesville, Virginia 22908, the ** Department of Orthopedic Surgery, University of Minnesota Cancer Center, Minneapolis, Minnesota 55455, the Dagger Dagger  Department of Urology and Winship Cancer Institute, Emory University, Atlanta, Georgia 30322, and the §§ Department of Urology, Indiana Cancer Pavilion, Indianapolis, Indiana 46202

Osteocalcin (OC) is a small (6 kDa) polypeptide whose expression was thought to be limited to mature osteoblasts. The discovery of OC expression in prostate cancer specimens led us to study the regulation of OC gene in androgen-independent metastatic human prostate PC3 cells. An 800-bp human OC (hOC) promoter-luciferase construct exhibited strong basal and vitamin D-induced activity in OC-positive human prostate and osteosarcoma cell lines. Through deletion analysis of the hOC promoter, the functional hierarchy of the cis-acting elements, OSE1, OSE2, and AP-1/VDRE, was established in PC3 cells (OSE1 > AP-1/VDRE > OSE2). By juxtaposing dimers of these 3 cis-elements, we produced a minimal hOC promoter capable of displaying high tissue specific activity in prostate cancer cells. Our study demonstrated three groups of transcription factors, Runx2, JunD/Fra-2, and Sp1, responsible for the high hOC promoter activity in PC3 cells by binding to the OSE2, AP-1/VDRE, and OSE1 elements, respectively. Among the three groups of transcription factors, the expression levels of Runx2 and Fra-2 are higher in the OC-positive PC3 cells and osteoblasts, compared with the OC-negative LNCaP cells. Interestingly, unlike the mouse OC promoter, the OSE1 site in hOC promoter is regulated by members of Sp1 family instead of the osteoblast-specific factor Osf1. The molecular basis for androgen-independent prostate cancer cells behaving like mature osteoblasts may be explained by the interplay and coordination of these transcription factors under the tight regulation of autocrine and paracrine mediators.

* This work was supported in part by National Institutes of Health Grant CA85555, the Kluge Foundation, DirectGene Inc., and NASA Grant NCC8-171 (to L. W. K. C.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¶¶ To whom correspondence should be addressed.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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