Elsevier

Matrix Biology

Volume 19, Issue 8, January 2001, Pages 717-725
Matrix Biology

Cysteine proteinases in chondrosarcomas

https://doi.org/10.1016/S0945-053X(00)00124-4Get rights and content

Abstract

The aim of the present study was to define the role of cathepsins B, H, K, L and S in the pathogenesis of human chondrosarcomas. For this purpose 40 tumour samples obtained from 12 patients with the diagnosis of conventional chondrosarcoma were systematically investigated for the expression of cathepsin mRNAs by Northern hybridisation, and for immunohistochemical localisation of the proteins. Northern analysis demonstrated the highest levels of cathepsins B and L in a recurring grade 1 chondrosarcoma, and in a grade 3 chondrosarcoma and in fibrous histiocytomas. Increased expression of cathepsin K mRNA was seen in seven chondrosarcomas, as well as in control tumours; fibrous histiocytomas, osteosarcomas, enchondromas and a giant cell tumour of bone. Cathepsin L was immunolocalised within the large chondrocytes, while cathepsin K was predominantly localised in large multinucleated osteoclastic cells and in some hypertrophic chondrocytes. These results suggest that chondrosarcoma can be included in the growing list of tumours, where cathepsins may well be involved in tumour progression. The simultaneous upregulation of cathepsins B and L, together with matrix metalloproteinase-13, and the association of cathepsin K with negative prognostic parameters suggests that an aggressive biological behaviour of chondrosarcoma may be related to the synthesis of cysteine proteinases and activation of other proteolytic enzymes. If this turns out to be the case, cathepsin inhibitors could provide the much needed adjuvant therapy for 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

References (32)

  • M.J. Bossard et al.

    Mechanism of inhibition of cathepsin K by potent, selective 1,5-diacylcarbohydrazides: a new class of mechanism-based inhibitors of thiol proteases

    Biochemistry

    (1999)
  • J.M. Chirgwin et al.

    Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease

    Biochemistry

    (1979)
  • B.D. Gelb et al.

    Pycnodysostosis, a lysosomal disease caused by cathepsin K deficiency

    Science

    (1996)
  • J.R. Griffiths

    Are cancer cells acidic?

    Br. J. Cancer

    (1991)
  • H.E. Gruber et al.

    Bone remodelling in the presence of chondrosarcoma: histomorphometry

    Acta Anat.

    (1993)
  • H.L. Evans et al.

    Prognostic factors in chondrosarcoma of bone

    Cancer

    (1977)
  • Cited by (30)

    • Cathepsins: Potent regulators in carcinogenesis

      2019, Pharmacology and Therapeutics
      Citation Excerpt :

      It has been identified as the principal protease in osteolytic lesions of giant cells in bone tumors (Lindeman et al., 2004). Expression of CTSK is upregulated in chondrosarcoma (Soderstrom, Ekfors, Bohling, Aho, & Vuorio, 2001), perivascular epithelioid cell tumors (PEComas) (Rao et al., 2013), and alveolar sarcoma (Zheng et al., 2013). In addition, its expression is entailed in the invasive growth of primary tumors in prostate cancer (Brubaker, Vessella, True, Thomas, & Corey, 2003), thyroid cancer (Mikosch et al., 2008), sub-cutaneous squamous cell carcinoma (Leusink et al., 2018), breast cancer, lung cancer, cervical cancer (Chen & Platt, 2011), renal cancer (Martignoni et al., 2012; Zheng et al., 2013), gastric carcinomas (Ren et al., 2012), melanoma (Rao et al., 2014), and glioblastoma multiforme (GBM) (Verbovsek et al., 2014).

    • Complexity of cancer protease biology: Cathepsin K expression and function in cancer progression

      2015, Seminars in Cancer Biology
      Citation Excerpt :

      CatK has been identified as the principal protease in osteolytic lesions of giant cells in bone tumours [100,101]. CatK was found to be upregulated in chondrosarcoma as well [102,103], where its expression levels correlated with metastatic potential and poor prognosis. CatK is expressed as well in renal and extrarenal perivascular epithelioid cell tumours (PEComas) [104,105] and in alveolar soft part sarcoma [106,107].

    • Circulating cathepsin K and cystatin C in patients with cancer related bone disease: Clinical and therapeutic implications

      2008, Biomedicine and Pharmacotherapy
      Citation Excerpt :

      As a consequence, any disregulation in the expression level of Cath K and/or Cyst C may result in disturbances of the normal metabolic turnover of bone tissue, which may lead to the onset of severe bone diseases. This hypothesis is corroborated by several clinical studies which highlighted that the expression levels of Cath K and/or Cyst C are altered in a number of nonmalignant and malignant diseases associated with an excessive bone resorption such as muscoloskeletal disorders, primary bone tumors or bone metastasis [4,8–18]. These findings suggest that Cath K and Cyst C may be regarded as suitable targets for appropriate treatments of these pathological processes and may be of value as additional circulating markers for the therapeutic monitoring and follow-up of patients with these diseases [4,9].

    • Cathepsin K is the principal protease in giant cell tumor of bone

      2004, American Journal of Pathology
      Citation Excerpt :

      A novel cathepsin K activity assay, based on the same principle as the MMP activity assays, was used to measure net cathepsin K activity. This assay revealed massive cathepsin K activity, which was more than 100-fold higher than activities found in other tissues expressing cathepsin K (ie, lung,23 atheroma,24 and chondrosarcoma25). No specific assays are as yet available for the other highly expressed cysteine protease cathepsin L. Traditional peptide-based activity assays lack specificity and are highly influenced by cathepsin K.

    • Cysteine proteases as disease markers

      2004, Clinica Chimica Acta
    View all citing articles on Scopus
    View full text