Abstract
Androgen refractory cancer of the prostate (ARCaP) cells contain androgen receptor (AR) and synthesize and secrete prostate specific antigen (PSA). We isolated epithelia-like ARCaPE from parental ARCaP cells and induced them to undergo epithelial–mesenchymal transition (EMT) by exposing these cells to soluble factors including TGFβ1 plus EGF, IGF-1, β2-microglobulin (β2-m), or a bone microenvironment. The molecular and behavioral characteristics of the resultant ARCaPM were characterized extensively in comparison to the parental ARCaPE cells. In addition to expressing mesenchymal biomarkers, ARCaPM gained 100% incidence of bone metastasis. ARCaPM cells express receptor activator of NF-κB ligand (RANKL), which was shown to increase tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts in culture, and when metastatic to bone in vivo. We provide evidence that RANKL expression was promoted by increased cell signaling mediated by the activation of Stat3-Snail-LIV-1. RANKL expressed by ARCaPM cells is functional both in vitro and in vivo. The lesson we learned from the ARCaP model of EMT is that activation of a specific cell signaling pathway by soluble factors can lead to increased bone turnover, mediated by enhanced RANKL expression by tumor cells, which is implicated in the high incidence of prostate cancer bone colonization. The ARCaP EMT model is highly attractive for developing new therapeutic agents to treat prostate cancer bone metastasis.
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Abbreviations
- AR:
-
Androgen receptor
- ARCaP:
-
Androgen-refractory human prostate cancer cell model from a patient with prostate cancer bone metastasis
- ARCaPE :
-
ARCaP clone with epithelial phenotype
- ARCaPM :
-
ARCaP clone with mesenchymal phenotype
- β2-m:
-
β2-Microglobulin
- BSP:
-
Bone sialoprotein
- C4-2:
-
Lineage derivative cells from LNCaP
- C4-2B:
-
C4-2 cells metastasized to bone
- CK18/19:
-
Cytokeratin 18/19
- CM:
-
Conditioned medium
- CREB:
-
cAMP-responsive element-binding protein
- E-cad:
-
E-cadherin
- EGF:
-
Epidermal growth factor
- EMT:
-
Epithelial–mesenchymal transition
- FBS:
-
Fetal bovine serum
- IC:
-
Intracardiac
- IGF-1:
-
Insulin-like growth factor 1
- IHC:
-
Immunohistochemistry
- IL13Rα2:
-
Interleukin13 receptor α2
- LNCaP:
-
Prostate cancer cells metastasized to lymph node
- MET:
-
Mesenchymal–epithelial transition
- MErT:
-
Mesenchymal–epithelial-reverting transition
- MMT:
-
Mesenchymal–mesenchymal transition
- N-cad:
-
N-cadherin
- NFκB:
-
Nuclear factor kappa B
- OC:
-
Osteocalcin
- OPG:
-
Osteoprotegerin
- OPN:
-
Osteopontin
- RANKL:
-
Receptor activator of NFκB ligand
- RT-PCR:
-
Reverse transcriptase-polymerase chain reaction
- SCID:
-
Severe combined immunodeficiency
- siRNA:
-
Small interfering RNA
- Stat:
-
Signal transducer and activator of transcription
- TGFβ1:
-
Transforming growth factor β1
- TRAP:
-
Tartrate-resistant acid phosphatase
- VM:
-
Vimentin
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Acknowledgements
We thank Gary Mawyer for editing. The helpful discussion and support from Guodong Zhu, Weiping Qian, Daqing Wu and Clayton Yates are greatly appreciated. The authors are particularly indebted to the helpful suggestions made by the editors, Drs. Rik Thompson and Elizabeth Williams, during the revision of this manuscript. This study was supported in part by CA082739 (HYEZ), U54 CA119338, CA098912 and CA766201 (LWKC). We are grateful to the generous gift from Frances and Clarence Wilkins in support of this study.
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Zhau, H.E., Odero-Marah, V., Lue, HW. et al. Epithelial to mesenchymal transition (EMT) in human prostate cancer: lessons learned from ARCaP model. Clin Exp Metastasis 25, 601–610 (2008). https://doi.org/10.1007/s10585-008-9183-1
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DOI: https://doi.org/10.1007/s10585-008-9183-1