Paracrine interactions between LNCaP prostate cancer cells and bioengineered bone in 3D in vitro culture reflect molecular changes during bone metastasis

Shirly Sieh; Anna V Taubenberger; Melanie L Lehman; Judith A Clements; Colleen C Nelson; Dietmar W Hutmacher

doi:10.1016/j.bone.2014.02.001

Paracrine interactions between LNCaP prostate cancer cells and bioengineered bone in 3D in vitro culture reflect molecular changes during bone metastasis

Bone. 2014 Jun:63:121-31. doi: 10.1016/j.bone.2014.02.001. Epub 2014 Feb 12.

Authors

Shirly Sieh¹, Anna V Taubenberger², Melanie L Lehman³, Judith A Clements⁴, Colleen C Nelson⁵, Dietmar W Hutmacher⁶

Affiliations

¹ Regenerative Medicine and Cancer Program, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland, Australia; Australian Prostate Cancer Research Centre-Queensland, Queensland University of Technology and Translational Research Institute, Brisbane, Queensland, Australia.
² Regenerative Medicine and Cancer Program, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland, Australia; BIOTEC TU Dresden, Dresden, Germany.
³ Australian Prostate Cancer Research Centre-Queensland, Queensland University of Technology and Translational Research Institute, Brisbane, Queensland, Australia; Department of Urologic Sciences, Vancouver Prostate Centre, University of British Columbia, Vancouver, British Columbia, Canada.
⁴ Cancer Program, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland, Australia.
⁵ Department of Urologic Sciences, Vancouver Prostate Centre, University of British Columbia, Vancouver, British Columbia, Canada; Cancer Program, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland, Australia.
⁶ Regenerative Medicine and Cancer Program, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland, Australia; Australian Prostate Cancer Research Centre-Queensland, Queensland University of Technology and Translational Research Institute, Brisbane, Queensland, Australia. Electronic address: dietmar.hutmacher@qut.edu.au.

PMID: 24530694
DOI: 10.1016/j.bone.2014.02.001

Abstract

As microenvironmental factors such as three-dimensionality and cell-matrix interactions are increasingly being acknowledged by cancer biologists, more complex 3D in vitro models are being developed to study tumorigenesis and cancer progression. To better understand the pathophysiology of bone metastasis, we have established and validated a 3D indirect co-culture model to investigate the paracrine interactions between prostate cancer (PCa) cells and human osteoblasts. Co-culture of the human PCa, LNCaP cells embedded within polyethylene glycol hydrogels with human osteoblasts in the form of a tissue engineered bone construct (TEB), resulted in reduced proliferation of LNCaP cells. LNCaP cells in both monoculture and co-culture were responsive to the androgen analog, R1881, as indicated by an increase in the expression (mRNA and/or protein induction) of androgen-regulated genes including prostate specific antigen and fatty acid synthase. Microarray gene expression analysis further revealed an up-regulation of bone markers and other genes associated with skeletal and vasculature development and a significant activation of transforming growth factor β1 downstream genes in LNCaP cells after co-culture with TEB. LNCaP cells co-cultured with TEB also unexpectedly showed similar changes in classical androgen-responsive genes under androgen-deprived conditions not seen in LNCaP monocultures. The molecular changes of LNCaP cells after co-culturing with TEBs suggest that osteoblasts exert a paracrine effect that may promote osteomimicry and modulate the expression of androgen-responsive genes in LNCaP cells. Taken together, we have presented a novel 3D in vitro model that allows the study of cellular and molecular changes occurring in PCa cells and osteoblasts that are relevant to metastatic colonization of bone. This unique in vitro model could also facilitate cancer biologists to dissect specific biological hypotheses via extensive genomic or proteomic assessments to further our understanding of the PCa-bone crosstalk.

Keywords: 3D cultures; Androgen; Bioengineered bone; Bone metastasis; Paracrine; Prostate cancer.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bone Neoplasms / etiology
Bone Neoplasms / metabolism*
Bone and Bones / cytology
Bone and Bones / metabolism
Cell Line, Tumor
Cell Proliferation / drug effects
Cells, Cultured
Coculture Techniques
Gene Expression Regulation, Neoplastic
Humans
Kallikreins / metabolism
Male
Metribolone / pharmacology
Paracrine Communication / drug effects
Polyethylene Glycols
Prostatic Neoplasms / complications
Prostatic Neoplasms / metabolism*
Tissue Engineering

Substances

Metribolone
Polyethylene Glycols
Kallikreins