Accession Number:

ADA582286

Title:

Prostatic Acid Phosphatase Plays a Causal Role in Prostate Cancer Bone Metastases

Descriptive Note:

Final rept. 1 Feb 2012-31 Jan 2013

Corporate Author:

MOUNT SINAI SCHOOL OF MEDICINE NEW YORK DEPT OF MEDICINE

Personal Author(s):

Report Date:

2013-02-01

Pagination or Media Count:

8.0

Abstract:

Bone metastases are the major cause of morbidity and mortality in prostate cancer PCa. While there are treatments for the osteolytic phase of PCa bone metastases, there are no therapies that inhibit the later, osteoblastic phase. Prostatic acid phosphatase PAP is a protein secreted by PCa cells and is highly expressed in PCa osteoblastic bone metastases. We previously demonstrated that PAP secreted by PCa cells induces the proliferation and differentiation of osteoblasts OB. We hypothesized that prostatic acid phosphatase PAP secreted by prostate cancer PCa cells in bone metastases plays a causal role in osteoblastic bone metastases. As the RANKRANKLOPG system coordinately controls the balance of osteoclasts vs. osteoblasts in bone, we determined the effects of PCa-derived PAP on RANKL and OPG expression in both PCa and OB cells. We demonstrate that PAP secretion by PCa cells modulates RANKLOPG secretion in both PCa and bone cells, favoring more OPG and less RANKL which would promote an osteoblastic phenotype. We utilized three different human PCa cell lines 1. VCaP PAP, induces osteoblastic lesions 2. PC3 PAP negative, induces osteolytic lesions and PC3ML more aggressive subline of PC3, also PAP negative and induces osteolytic lesions. We successfully generated cell lines that transiently knockdown PAP expression using RNAi technology PAP-siRNA in high PAP-expressing human PCa cells VCaP-luc. Conversely, for the gain-of-function experiments, PAP-negative human PCa cells PC3-luc and a more aggressive subline PC3-ML-luc were stably transfected to overexpress secretory PAP by utilizing the retroviral expression vector pLenti-GIII-CMV-hACPP ACPP is PAP Lentiviral Vector PC3-pPAP and the pLenti-III-HA Blank Control PC3-luc-control vector as control. Stable cell clones were selected with puromycin and characterized by qRT-PCR and western blotting. Knockout of PAP in VCaP cells induced cell cycle arrest. Knock-in of PAP in PC3 and PC3ML

Subject Categories:

  • Biochemistry
  • Anatomy and Physiology
  • Medicine and Medical Research

Distribution Statement:

APPROVED FOR PUBLIC RELEASE