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Establishment and validation of an in vitro co-culture model to study the interactions between bone and prostate cancer cells

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Abstract

Bone is the preferred site for prostate cancer (PCa) metastases. Once the tumor has established itself within the bone there is virtually no cure. To better understand the interactions between the PCa cells and bone environment in the metastatic process new model systems are needed. We have established a two-compartment in vitro co-culturing model that can be used to follow the trans-activation of bone and/or tumor cells. The model was validated using two PCa tumor cell lines (PC-3; lytic and LNCaP; mixed/osteoblastic) and one osteolytic inducing factor, 1,25-dihydroxyvitamin D3 (D3). Results were in accordance with the expected bone phenotypes; PC-3 cells and D3 gave osteolytic gene expression profiles in calvariae, with up-regulation of genes needed for osteoclast differentiation, activation and function; Rankl, CathK, Trap and MMP-9, and down-regulation of genes associated with osteoblast differentiation and bone mineralization; Alp, Ocl and Dkk-1. LNCaP cells activated genes in the calvarial bones associated with osteoblast differentiation and mineralization, with marginal effects on osteolytic genes. The results were strengthened by similar changes in protein expression for a selection of the analyzed genes. Furthermore, the osteolytic gene expression profiles in calvarial bones co-cultured with PC-3 cells or with D3 were correlated with the actual ongoing resorptive process, as assessed by the release of collagen fragments from the calvariae. Our results show that the model can be used to follow tumor-induced bone remodeling, and by measuring changes in gene expression in the tumor cells we can also study how they respond to the bone microenvironment.

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Abbreviations

ACTB:

β-Actin

ALP:

Alkaline phosphatase

CATHK:

Cathepsin K

CTX-I:

C-terminal telopeptide fragments of type I collagen

D3:

1,25-Dihydroxyvitamin D3

DKK-1:

Dickkopf-1

ET-1:

Endothelin 1

ETA:

Endothelin receptor A

IGF-1:

Insulin-like growth factor type 1

IGF-R1:

Insulin-like growth factor type 1 receptor

MMP-9:

Matrix metalloproteinase 9

OCL:

Osteocalcin

OPG:

Osteoprotegerin

PCa:

Prostate cancer

PCR:

Polymerase chain reaction

PSA:

Prostate specific antigen

PTHRP:

Parathyroid hormone-related protein

RANK:

Receptor activator of nuclear factor-κB

RANKL:

Receptor activator of nuclear factor-κB ligand

RPL13:

Ribosomal protein L13

TGF-β1:

Transforming growth factor β 1

TRAP:

Tartrate-resistant acid phosphatase

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Acknowledgments

This work was supported by grants from the Swedish Cancer Society (project no. 070604), the Swedish Medical Research Council (project no. 7525), The Royal 80 Year Fund of King Gustav V, the Norwegian Cancer Society, Lions Research Foundation and Umeå University, Sweden.

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Authors and Affiliations

  1. Department of Radiation Sciences, Oncology, Umeå University, 901 85, Umeå, Sweden

    Annika Nordstrand, Jonas Nilsson, Åse Tieva & Anders Widmark

  2. Department of Medical Biosciences, Pathology, Umeå University, Umeå, Sweden

    Pernilla Wikström

  3. Department of Oral Cell Biology, Umeå University, Umeå, Sweden

    Ulf H. Lerner

Authors
  1. Annika Nordstrand
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  2. Jonas Nilsson
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  3. Åse Tieva
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  4. Pernilla Wikström
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  5. Ulf H. Lerner
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  6. Anders Widmark
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Corresponding author

Correspondence to Annika Nordstrand.

Additional information

Annika Nordstrand and Jonas Nilsson contributed equally to this work.

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Nordstrand, A., Nilsson, J., Tieva, Å. et al. Establishment and validation of an in vitro co-culture model to study the interactions between bone and prostate cancer cells. Clin Exp Metastasis 26, 945–953 (2009). https://doi.org/10.1007/s10585-009-9285-4

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  • DOI: https://doi.org/10.1007/s10585-009-9285-4

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