Prostate epithelial cell lines form spheroids with evidence of glandular differentiation in three-dimensional Matrigel cultures

Abstract

Normal (PNT2-C2) and metastatic (PC-3) prostate cell lines were grown in Matrigel to observe the effects on morphology and phenotype in comparison to monolayer culture. In monolayer cultures, PNT2-C2 showed typical round/cuboidal epithelial morphology, with tight cell associations, whereas in Matrigel they formed smooth spheroids, tightly packed with cells. In both monolayer and Matrigel, PNT2-C2 had a differentiated luminal epithelial phenotype with high expression of cytokeratin 8, prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), E-cadherin and desmoglein. In contrast, PC-3 cells possessed an epithelial/mesenchyme morphology in monolayer with loose cell to cell contact and pseudopodial extensions. Immunohistochemical phenotyping indicated the cells were undifferentiated, expressing high levels of vimentin, β1 integrin, CD44 and low expression of cytokeratin 8. In Matrigel they formed smooth and irregular spheroids, which had a lumen surrounded by a single cell layer. Matrigel also influenced the expression of PSA, PSMA and CD44. These results indicate that Matrigel culture can induce morphological differentiation of prostate cancer cells which initially had a basal phenotype. © 2001 CancerResearch Campaign http://www.bjcancer.com

Incorporation of a laminin-derived peptide (SIKVAV) on polymer-modified adenovirus permits tumor-specific targeting via α6-integrins

“…Importantly, the retargeted virus demonstrated no transduction above that of pc-Adluc, in the cells derived from normal prostate epithelia suggesting that the risk of inappropriate transgene expression in healthy tissue would be low. Although we detected b1 integrin subunits on both PNT1a and PNT2 cells in agreement with previous studies, 50,51 we would expect these cells to maintain b4 integrin subunit expression to which the a6 subunit would preferentially dimerize, SIKVAV being unable to bind integrin a6b4. 52 Summary of flow cytometric analysis using specific monoclonal anti-a6-integrin subunit and monoclonal anti-b1-integrin subunit antibodies on a panel of human cell lines.…”

BMP7, a Putative Regulator of Epithelial Homeostasis in the Human Prostate, Is a Potent Inhibitor of Prostate Cancer Bone Metastasis in Vivo

“…[57][58][59] The observed reversion to an epithelial morphology in prostate cancer seems possible even at bone metastatic sites [57][58][59] and is in full agreement with our in vitro and in vivo data presented here. Furthermore, previous studies demonstrated cellular plasticity of human prostate cancer cells (PC3 cells), particularly the EMT/MET interconversion induced by different culture conditions (monolayer versus three-dimensional cultures 60 ). In many prostate cancer patients, micrometastatic deposits may already exist in bone marrow after removal of the primary tumor that cannot be identified at the time of diagnosis (minimal residual disease).…”

Isolation and functional characterization of murine prostate stem cells

“…Assays developed to measure neural and mammary stem cell self-renewal in vitro propagate these cells as spheres in an attempt to preserve their specialized biology (23,24). Several studies have shown that human prostate cells can form epithelial spheroids when cocultured with stromal cells and suspended in Matrigel in vitro (25)(26)(27). This strategy was adapted to develop a method for measuring murine prostate stem cell self-renewal.…”