BERKELEY, CA (UroToday.com) - To date, the majority of in vitro studies carried out in prostate cancer (PCa) have been undertaken using traditional 2D models. In these models, many properties found in vivo are lost, including cell-cell communication, polarisation, and extracellular contacts, while wound healing, inflammation responses, and proliferation are artificially promoted, well above the normal rates for in vivo situations.[1, 2] Alternatively, 3D cell culture mimics the in vivo environment more realistically than 2D monolayers, allowing for the subtle interplay of cells of the same or different origins within a support network. The importance of studying cancer cells in 3D culture has been demonstrated previously, where in comparison to 3D, 2D cultures failed to generate invasive or metastatic sublines.[2] Moreover, proper alignment and spatial organisation of cells in 3D is essential for tumour progression.[2] Three dimensional models which assess the microenvironment and cellular behaviour are, therefore, becoming essential tools for exploring metastasis.
Antigenic profiles of tumours excised from advanced PCa patients have identified alterations in the expression of numerous proteins. Of these, the androgen receptor (AR),[3] α6 integrin [4, 5] and β1 integrin subunit,[6] vimentin,[7] cell adhesion molecules E Cadherin and N Cadherin,[8] and, more recently, chemokine receptor CXCR4 [9] expression have been linked to increased Gleason grade and metastatic dissemination in PCa. Using these well-established markers, we present novel data concerning the direct differences in protein expression exhibited by a range of PCa cell-lines using both traditional 2D monolayer vs 3D culture systems.
Utilising Matrigel™ we cultured a range of PCa cell-lines including non-cancerous (RWPE-1), non invasive (LNCaP), and metastatic (PC3, DU145) cells in 3D over a 9-day period. In contrast to 2D monolayers, when plated in 3D, these cells displayed distinct differences in cell morphology, proliferation, and expression of important biomarker proteins associated with cancer progression. For example, consistent with protein expression patterns of metastatic tumour formation as found in vivo, metastatic PC3 cell-lines exhibited a down-regulation of E-cadherin and α6 integrin expression and an up-regulation of N-cadherin, vimentin and β1 integrin expression and re-expressed non-transcriptionally active AR. In 2D cultures, there was little distinction in protein expression between metastatic, non-invasive and epithelial cells. Our results clearly indicate that 3D cultures afford a better platform that discerns antigenic profiles, which more directly mimic that found in vivo.
References:
- Harma, V., J. Virtanen, R. Makela, A. Happonen, J.P. Mpindi, M. Knuuttila, P. Kohonen, J. Lotjonen, O. Kallioniemi, and M. Nees, A comprehensive panel of three-dimensional models for studies of prostate cancer growth, invasion and drug responses. PLoS One, 2010. 5(5): p. e10431.
- Nyga, A., U. Cheema, and M. Loizidou, 3D tumour models: novel in vitro approaches to cancer studies. J Cell Commun Signal, 2011.
- Tamburrino, L., F. Salvianti, S. Marchiani, P. Pinzani, G. Nesi, S. Serni, G. Forti, and E. Baldi, Androgen receptor (AR) expression in prostate cancer and progression of the tumor: Lessons from cell lines, animal models and human specimens. Steroids 2012.
- Knox, J.D., A.E. Cress, V. Clark, L. Manriquez, K.S. Affinito, B.L. Dalkin, and R.B. Nagle, Differential expression of extracellular matrix molecules and the alpha 6-integrins in the normal and neoplastic prostate. Am J Pathol, 1994. 145(1): p. 167-74.
- Bonkhoff, H., U. Stein, and K. Remberger, Differential expression of alpha 6 and alpha 2 very late antigen integrins in the normal, hyperplastic, and neoplastic prostate: simultaneous demonstration of cell surface receptors and their extracellular ligands. Hum Pathol, 1993. 24(3): p. 243-8.
- Murant, S.J., J. Handley, M. Stower, N. Reid, O. Cussenot, and N.J. Maitland, Co-ordinated changes in expression of cell adhesion molecules in prostate cancer. Eur J Cancer, 1997. 33(2): p. 263-71.
- Lang, S.H., C. Hyde, I.N. Reid, I.S. Hitchcock, C.A. Hart, A.A. Bryden, J.M. Villette, M.J. Stower, and N.J. Maitland, Enhanced expression of vimentin in motile prostate cell lines and in poorly differentiated and metastatic prostate carcinoma. Prostate, 2002. 52(4): p. 253-63.
- Wang, J., D. Krill, M. Torbenson, Q. Wang, M. Bisceglia, J. Stoner, A. Thomas, P. DeFlavia, R. Dhir, and M.J. Becich, Expression of cadherins and catenins in paired tumor and non-neoplastic primary prostate cultures and corresponding prostatectomy specimens. Urol Res, 2000. 28(5): p. 308-15.
- Wang, J., R. Loberg, and R.S. Taichman, The pivotal role of CXCL12 (SDF-1)/CXCR4 axis in bone metastasis. Cancer Metastasis Rev, 2006. 25(4): p. 573-87.
Written by:
Louisa C.E. Windus as part of Beyond the Abstract on UroToday.com. This initiative offers a method of publishing for the professional urology community. Authors are given an opportunity to expand on the circumstances, limitations etc... of their research by referencing the published abstract.
Eskitis Institute for Cell and Molecular Therapies
Discovery Biology
Griffith University
Nathan 4111
Brisbane, Queensland, Australia
In vivo biomarker expression patterns are preserved in 3D cultures of prostate cancer - Abstract
More Information about Beyond the Abstract