Expression of Cathepsin B and L antigen and activity is associated with early colorectal cancer progression

https://doi.org/10.1016/j.ejca.2004.03.011Get rights and content

Abstract

Cathepsin B and Cathepsin L are cysteine proteases important in the process of invasion and metastasis. The aim of our study was to assay antigen and activity levels of these enzymes and to correlate these with established clinical and pathological prognostic parameters including patient survival. 99 patients undergoing operations for colorectal cancer were included in this study. We quantitated cathepsin B and L levels in matched normal mucosa and cancer samples using an enzyme-linked immunosorbent assay (ELISA) and specific activity assays and expressed the results as tumour/normal ratios. Significant correlations were found between tumour/normal cathepsin B and L antigen and activity ratios. Cathepsin B and L tumour/normal activity ratios were greater than 1 in early stage disease and there were gradual reductions in cathepsin B (P=0.02) and L (P=0.03) activity ratios with advancing tumour stage. Survival of patients with potentially curative disease was inversely related to both cathepsin B (P=0.007) and L (P=0.001) activity ratio, in addition to cathepsin L antigen ratio (P=0.008). Our findings suggest that cysteine proteases play an important role in colorectal cancer progression.

Introduction

The malignancy of solid tumours is largely related to their capacity for invasion and metastasis, and proteases may play a major role in this process. The lysosomal cysteine protease, cathepsin B, found in both tumour epithelium and stroma [1], is known to degrade components of the extracellular matrix and basement membrane [2] and has been measured in both primary and metastatic colorectal cancers [3]. It converts inactive pro-urokinase-type plasminogen activator (uPA) to active uPA [4], and uPA and its inhibitor plasminogen activator inhibitor 1 (PAI-1) have been shown to be closely associated with outcome in many tumour types including colorectal, breast, gastric and lung cancers [5]. Levels of both the active and precursor forms of cathepsin B are elevated compared with normal tissue in many cancer types, as well as in transformed cells in culture [6], [7], [8], [9] and the specific activity of cathepsin B has been found to correlate with tumorigenicity in a number of cell lines [10]. Cathepsin L levels have also been shown to be elevated both in cancer cell lines and in colorectal tumours [8], [9], [11], [12], [13].

Although cathepsin B and L have previously been correlated with clinico-pathological features in a number of malignant tumours [5], information on their prognostic value is limited in relation to colorectal cancer. In this study, we aimed to correlate cathepsin B and L activity and antigen levels with clinical and pathological features and with cancer-specific survival in a large cohort of colorectal cancer patients.

Section snippets

Patients and tissue collection

Ninety nine patients (median age 71 years, range 31–84 years; 59 male) undergoing colorectal cancer surgery in St Vincent's University Hospital, Dublin between September 1991 and December 1994 were included in this prospective study which was approved by the hospital's ethics committee. For the purposes of this study, colorectal carcinoma tissue and normal colorectal mucosa (macroscopically normal at least 5cm away from the tumour margin) from the same patient was resected immediately following

Cathepsin antigen and activity expression in normal and cancer tissues

Cathepsin B antigen ranged from 0.09 to 60.0 ng/mg protein in normal mucosa and from 0.02 to 104.0 ng/mg protein in colorectal cancer. Cathepsin B activity varied from 0.013 to 3.4 U/mg protein in normal tissue and from 0.007 to 20.0 U/mg protein in cancer tissue. Cathepsin L antigen expression in normal mucosa and cancer tissues ranged from 0.20 to 17.0 and from 0.02 to 60.7 ng/mg protein, respectively. Cathepsin L activity ranged from 0.02 to 4.8 U/mg protein in normal tissue and from 0.01 to

Discussion

The breakdown of natural barriers to tumour spread is an essential component of invasion and metastasis. A number of proteolytic enzymes have been implicated in this process including serine, aspartyl and matrix metalloproteases. Cysteine proteases, such as cathepsin B and L, are also believed to play a central role by catalysing the degradation of several components of the extracellular matrix. Cathepsin B and L have previously been found to correlate with metastatic potential in a number of

Acknowledgments

This work was funded by a grant from the Irish Cancer Society. The authors would like to acknowledge the assistance of the Departments of Surgery and Histopathology, St. Vincent's University Hospital and Ms. Anne White.

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