Hyaluronan (HA) interacting proteins RHAMM and hyaluronidase impact prostate cancer cell behavior and invadopodia formation in 3D HA-based hydrogels

• Published in: PloS one Vol. 7; no. 11; p. e50075
• Main Authors: Gurski, Lisa A, Xu, Xian, Labrada, Lyana N, Nguyen, Ngoc T, Xiao, Longxi, van Golen, Kenneth L, Jia, Xinqiao, Farach-Carson, Mary C
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 16.11.2012; Public Library of Science (PLoS)
• Subjects: Acids; B cells; Biocompatibility; Bioengineering; Biological activity; Biology; Biotinylation; Blotting, Western; Bone marrow; Cancer; Cancer metastasis; CD44 antigen; Cell adhesion & migration; Cell culture; Cell growth; Cell Migration Assays - instrumentation; Cell Migration Assays - methods; Cell Surface Extensions - physiology; Clusters; Connective tissues; Crosslinking; Dissection; Extracellular Matrix Proteins - genetics; Extracellular Matrix Proteins - metabolism; Gene Knockdown Techniques; Growth factors; Humans; Hyaluronan Receptors - genetics; Hyaluronan Receptors - metabolism; Hyaluronic acid; Hyaluronic Acid - metabolism; Hyaluronoglucosaminidase - metabolism; Hydrogels; Hydrogels - metabolism; Invasiveness; Kinases; Male; Materials Science; Medical prognosis; Medical research; Medicine; Metastases; Metastasis; Molecular weight; Motility; Neoplasm Invasiveness - physiopathology; Prostate cancer; Prostatic Neoplasms - metabolism; Prostatic Neoplasms - physiopathology; Proteins; Rheology; Tissues; Trypan Blue; Tumor Cells, Cultured; Delaware; United States > US
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0050075

To study the individual functions of hyaluronan interacting proteins in prostate cancer (PCa) motility through connective tissues, we developed a novel three-dimensional (3D) hyaluronic acid (HA) hydrogel assay that provides a flexible, quantifiable, and physiologically relevant alternative to current methods. Invasion in this system reflects the prevalence of HA in connective tissues and its role in the promotion of cancer cell motility and tissue invasion, making the system ideal to study invasion through bone marrow or other HA-rich connective tissues. The bio-compatible cross-linking process we used allows for direct encapsulation of cancer cells within the gel where they adopt a distinct, cluster-like morphology. Metastatic PCa cells in these hydrogels develop fingerlike structures, "invadopodia", consistent with their invasive properties. The number of invadopodia, as well as cluster size, shape, and convergence, can provide a quantifiable measure of invasive potential. Among candidate hyaluronan interacting proteins that could be responsible for the behavior we observed, we found that culture in the HA hydrogel triggers invasive PCa cells to differentially express and localize receptor for hyaluronan mediated motility (RHAMM)/CD168 which, in the absence of CD44, appears to contribute to PCa motility and invasion by interacting with the HA hydrogel components. PCa cell invasion through the HA hydrogel also was found to depend on the activity of hyaluronidases. Studies shown here reveal that while hyaluronidase activity is necessary for invadopodia and inter-connecting cluster formation, activity alone is not sufficient for acquisition of invasiveness to occur. We therefore suggest that development of invasive behavior in 3D HA-based systems requires development of additional cellular features, such as activation of motility associated pathways that regulate formation of invadopodia. Thus, we report development of a 3D system amenable to dissection of biological processes associated with cancer cell motility through HA-rich connective tissues.