Anti-tumor activities of chondroitinase AC and chondroitinase B: inhibition of angiogenesis, proliferation and invasion

doi:10.1016/S0014-2999(01)00884-6

European Journal of Pharmacology

Volume 416, Issue 3, 30 March 2001, Pages 213-221

https://doi.org/10.1016/S0014-2999(01)00884-6 Get rights and content

Abstract

In the current study, two specific glycosaminoglycan lyases, chondroitinase AC and chondroitinase B, were utilized to examine the roles of chondroitin sulfates and dermatan sulfate in tumor metastasis and angiogenesis. Melanoma cells (SK-MEL) or endothelial cells were treated with either medium or chondroitinase enzyme. Chondroitinase AC inhibited melanoma invasion and proliferation as well as endothelial proliferation and angiogenesis. Apoptosis of melanoma and endothelial cells, as measured by the activity of caspase-3, was also increased by chondroitinase AC, but not by chondroitinase B. Chondroitinase B inhibited endothelial and melanoma proliferation and invasion, but to a lesser extent than chondroitinase AC. Neither chondroitinase had a detectable effect on gelatinase secretion by melanoma cells. These results indicate that both chondroitin and dermatan sulfates regulate many cellular activities related to metastasis.

Introduction

The growth of tumors requires tumor cell proliferation as well as the formation of new blood vessels. Metastasis of the primary tumor and growth of secondary tumors involve a complex sequence of events, which has been referred to as the “metastatic cascade” (Sneath and Mangham, 1998). While many growth factors, enzymes and extracellular matrix components are involved in these events, works by other investigators have suggested that chondroitin sulfate proteoglycans may play a key role in tumor growth and metastasis.

In examining the functions of chondroitin sulfates in metastasis, two different approaches have been used: inhibiting the formation of chondroitin sulfate proteoglycans with β-d-xyloside (ρ-nitrophenyl-β-d-xylopyranoside), and enzymatically removing chondroitin sulfates from cells with chondroitinase ABC. Both methods have similar results and effectively decrease tumor cell invasion and endothelial migration and adhesion Henke et al., 1996, Faassen et al., 1992, Faassen et al., 1993.

Taken together, studies utilizing inhibitors or enzymes, which degrade chondroitin sulfates, indicate that chondroitin sulfates play important roles in the processes of tumor growth and metastasis Iida et al., 1996, Sneath and Mangham, 1998. However, it is not clear from these prior studies which chondroitin sulfate proteoglycans were involved. Chondroitin sulfate proteoglycans exist in three major forms: chondroitin sulfate A, chondroitin sulfate B (dermatan sulfate) and chondroitin sulfate C. Recent work has demonstrated that the different forms of chondroitin sulfates have different functions. Dermatan sulfate is important in controlling fibroblast proliferation, and chondroitin sulfates A and C regulate integrin-mediated cell adhesion Denholm et al., 2000, Moyano et al., 1999.

The purpose of the present study was to determine the relative importance of the different forms of chondroitin sulfate in cellular activities related to metastasis. To accomplish this, two specific glycosaminoglycan lyases were utilized: chondroitinase AC (substrates are chondroitin sulfates A and C) and chondroitinase B (substrate is chondroitin sulfate B or dermatan sulfate). The effects of these two glycosaminoglycan lyases on several tumor and endothelial cell functions were compared, and the mechanisms underlying the inhibition of these activities were examined.

Section snippets

Materials

Human melanoma (SK-MEL-2) and calf pulmonary artery endothelial cells were from ATCC, Manassas, VA. Dulbecco's minimal essential medium, Fischer's medium, phosphate buffered saline (PBS) and fetal bovine serum were from Gibco, Grand Island, NY. Dermatan sulfate, chondroitin sulfates A and C, and heparan sulfate were purchased form Celsus Laboratories, Cincinnati, OH. Basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) were purchased from Peprotech, Rocky Hill,

Angiogenesis and endothelial cell proliferation

Capillary endothelial cells were treated with 0.1–10 IU/ml of chondroitinase AC or chondroitinase B, and angiogenesis, as measured by the formation of capillary-like structures, was examined. Endothelial cells treated with chondroitinase AC contained fewer capillary-like structures than untreated controls did Fig. 1, Fig. 2. Angiogenesis was inhibited by 46% and 72%, following treatment with 1.0 and 10 IU/ml of chondroitinase AC, respectively. In comparison, β-d-xyloside, which blocks the

Discussion

Comparison of the relative effects of chondroitinase AC with those of chondroitinase B, indicated that both chondroitin and dermatan sulfates are involved in regulating a number of cellular activities linked to metastasis. At the start of these experiments, it had been thought that the use of the two specific chondroitinase enzymes would reveal a clearly predominant role for either chondroitin or dermatan sulfates in each of these various activities. It would appear that chondroitin sulfates A

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Anti-tumor activities of chondroitinase AC and chondroitinase B: inhibition of angiogenesis, proliferation and invasion

Abstract

Introduction

Section snippets

Materials

Angiogenesis and endothelial cell proliferation

Discussion

Eur. J. Pharmacol.

Biochim. Biophys. Acta

J. Biol. Chem.

Semin. Cancer Biol.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Eur. J. Pharmacol.

Carbohydr. Res.

Impact of various glycosaminoglycans upon cAMP-dependent protein kinase

Cell Biochem. Funct.

Melanoma chondroitin sulphate proteoglycan regulates cell spreading through Cdc42, Ack-1 and p130cas

Nat. Cell Biol.

Cell surface chondroitin sulfate proteoglycan, immunologically related to CD44, is involved in Type I collagen-mediated melanoma motility and invasion

J. Cell Biol.

Melanoma chondroitin sulphate proteoglycan regulates cell spreading through Cdc42, Ack-1 and p130^cas