Rabbit Anti-Human HYAL1 Polyclonal antibody

Interactions of hyaluronan (HA) and tumor and stromal cells are highly discussed as one of the major contributors in tumor progression and metastasis. The balance of HA in the tissue is highly regulated by two key enzyme classes; hyaluronan synthases (HAS) and hyaluronidases (HYAL). Current reports hint that the HA amount in the tissue is correlated with poor prognosis in melanoma, the most life-threatening skin tumor. In this work, we generated in vivo mouse models with low and high expression of Has2 and used the models for studying melanoma proliferation of the B78D14 melanoma cell line. We found that a strong reduction of HA amount in the skin was correlated to decreased tissue stiffness and a reduction in tumor weight. Since tumor cells have a direct contact to the HA in the tumor and at the stroma interface, we reconstituted different biomimetic in vitro models using fibroblasts derived from a mouse model to recapitulate melanoma cell behavior at the tumor boundary, namely, (i) decellularized fibroblast matrix (FbECM), (ii) fibroblast embedded into 3D collagen matrices (FbColl), and (iii) well-defined HA-functionalized 3D collagen matrices (HAColl). We found no considerable effect of high and low amounts of fibroblast-derived HA in the matrices on melanoma proliferation and invasion. However, HYAL1-treated FbECM and FbColl, as well as HAColl functionalized with low molecular weight HA (34 kDa) promoted proliferation and invasion of melanoma cells in a concentration dependent manner. Our results emphasize the molecular weight specific effects of HA in regulation of melanoma behavior and provide an alternative explanation for the in vivo observation of HA dependent tumor growth.

Background: Hyaluronan is a major component of the extracellular matrix of periodontal ligament (PDL) contributing to the structural and functional integrity. Hyaluronans contribute to the buffering effect of the PDL during chewing, and they are also important in inflammation and wound healing. Hyaluronan is known to be synthesized and turned over by the resident PDL cells, although the mechanisms of hyaluronan metabolism still remain unclear. Hyaluronidase (HAase), an endoglycosidase, degrades hyaluronan into small fragments. Currently, 3 human HAases, HYAL1, HYAL2, and PH-20, have been identified and well characterized. Methods: This study was conducted to investigate the expression and activity of these HAases in cultured human PDL fibroblasts and to elucidate the mechanisms involved in hyaluronan metabolism under normal and inflammatory conditions. Human PDL fibroblasts derived from the periodontium of 3 premolars were cultured with or without interleukin (IL)-1beta (0.1 to 10 ng/ml) and tumor necrosis factor (TNF)-alpha (1 to 100 ng/ml) for 0 to 48 hours. The expression of.HAase mRNA was assessed by reverse transcription-polymerase chain reaction (RT-PCR) and quantitative real-time PCR, and the enzymatic activity was examined using hyaluronan zymography. Results: PDL fibroblasts expressed HYAL1 and HYAL2 mRNAs, but not PH-20 mRNA. The expression of HYAL1 mRNA was enhanced by about 3.5- and 3.7-fold at maximum after 1 hour stimulation with 1 ng/ml ILAP and after 3-hour stimulation with 10 ng/ml TNF-alpha, respectively. The expression of HYAL2 and PH-20 mRNAs was not affected by stimulation with cytokines. HAase activity was detected in conditioned medium from PDL fibroblast cultures, and the activity was enhanced by treatment with 10 ng/ml TNF-alpha. Conclusion: These results suggest that PDL fibroblasts express HAases and generate HAase activity essential for extracellular hyaluronan metabolism under physiological and inflammatory conditions.