Cysteine proteinases in chondrosarcomas
Introduction
Degradation of extracellular matrix is essential for tumour proliferation, growth and metastasis. Cysteine proteinases are a family of enzymes, which together with matrix metalloproteinases are involved in the destruction of both collagen and proteoglycan components of the extracellular matrix (Maciewicz and Wotton, 1991). This includes both physiologic processes such as endochondral ossification and bone remodelling, and a variety of pathological events such as tumour invasion and growth. Cathepsins B and L have been implicated in the development of several gastrointestinal and some other tumours through the destruction of surrounding extracellular matrix components (Ohta et al., 1994, Herszényi et al., 1999), but there is also evidence for activation of cathepsins H and S in malignant tumours (Kirschke et al., 1998). Osteoclastoma-derived cell lines have been demonstrated to overproduce cathepsin K (Li et al., 1995). Furthermore, another cysteine cathepsin, named cathepsin O, has been cloned from a breast carcinoma (Velasco et al., 1994). The increases in cysteine proteinase activity in malignant tumours may reflect alterations in the synthesis, activation and/or intracellular processing pathways of the enzymes. In addition, the increased activity of these proteinases may be due to a decrease in the levels of specific cysteine proteinase inhibitors, cystatins (Bobek and Levine, 1992).
Chondrosarcoma is the second most common primary malignant bone tumour, originating from cartilage cells. Primary chondrosarcoma arises de novo, and has the ability to degrade and invade normal bone tissue. Typical features of chondrosarcomas are their inconspicuous mode of growth, lack of symptoms and often a large size at the time of primary diagnosis (Unni, 1996). The tumour typically expands within bone, but also grows through the cortex. The mechanisms involved in the expansion of chondrosarcomas remain poorly understood; stimulation of osteoclastic bone resorption has been suggested (Gruber et al., 1993). Pathologic classification of chondrosarcomas into grades 1, 2 and 3 tumours has traditionally been used to assess the treatment principles and clinical outcome of patients with chondrosarcoma (Evans et al., 1977). However, histologic interpretation and prediction of biologic behaviour of this tumour is often extremely difficult. The need for an objective immunohistochemical marker predicting the survival of chondrosarcoma patients is obvious.
Surprisingly, essentially no information is available about the role of cysteine proteinases in chondrosarcomas. This, and the finding that cathepsin K is a major product of osteoclasts and some hypertrophying chondrocytes (Li et al., 1995, Rantakokko et al., 1996) prompted us to examine the possibility that cysteine cathepsins could also be associated with invasion and growth of malignant cartilaginous tumours. Defining the role of cysteine proteinases in the invasiveness of chondrosarcomas is important in order to understand the biology of tumour invasion and growth, and to search for new diagnostic tools, prognostic indicators and therapeutic targets. The purpose of the present study was to examine the expression of cysteine proteinases at mRNA level and to perform their immunohistochemical localisation in human chondrosarcomas, in eight control tumours and in cartilage and bone.
Section snippets
Patients
The material consists of 40 tumour samples obtained from 12 patients with the diagnosis of conventional chondrosarcoma. The samples were obtained from the Department of Surgery, Turku University Central Hospital and from the Department of Orthopaedics and Traumatology, Helsinki University Central Hospital. Table 1 shows the clinical characteristics of the tumours. All patients were treated primarily by surgery. In addition, samples from two osteosarcomas, two fibrotic histiocytomas, one giant
Expression of cathepsin mRNAs
The expression of cathepsin genes in human chondrosarcomas was studied by Northern analysis of total RNAs from the different tumour specimens (Fig. 1). The hybridisation signals were quantified and corrected for variations in RNA loading, determined by the level of 28S rRNA (Fig. 2). Two distinct cathepsin B mRNAs of 2.2 and 4.0 kb were detected in most tumour samples (Fig. 1). The highest levels of cathepsin B mRNA were seen in chondrosarcomas #7 (a recurring grade 1 tumour) and #11 (grade 3),
Discussion
The ability of tumour cells to invade tissues by degrading extracellular matrix is characteristic for malignant neoplasms. Several studies are available on the prognostic relevance of cysteine proteinases in tumours of mesenchymal and epithelial origin (Otto et al., 1999, Yan et al., 2000), but none have evaluated the role of cathepsins in chondrosarcomas. Our study demonstrates for the first time the presence of cathepsins B, H, K, L and S mRNAs in human chondrosarcomas, and suggests that at
Acknowledgements
We are grateful to Dr. Dieter Brömme for the cathepsin K antibody and to Dr. Heidrun Kirschke for the antibodies against cathepsins B, H, L and S. Dr. Sakamuri Reddy is acknowledged for generously providing the cDNA probe for mouse TRAP. The expert technical assistance of Päivi Auho, Merja Lakkisto and Tuula Oivanen is gratefully acknowledged. This study was financially supported by the Academy of Finland (project no 37311), the University Central Hospital of Turku (project no.s 13304 and
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