Search tips
Search criteria 


Logo of brjcancerBJC HomepageBJC Advance online publicationBJC Current IssueSubmitting an article to BJCWeb feeds
Br J Cancer. 1999 October; 81(3): 510–519.
PMCID: PMC2362911

Immunochemical analysis of cathepsin B in lung tumours: an independent prognostic factor for squamous cell carcinoma patients


In order to evaluate the possible role of the proteolytic enzyme cathepsin B (cath B) in human non-small cell lung cancer (NSCLC) we examined cath B concentrations (cath Bc) and activities (cath BA) in homogenates of 127 pairs of lung tumour tissues and corresponding non-tumourous lung parenchyma. Total cath B activity (cath BAT) and enzymatic activity of the fraction of cath B, which is stable and active at pH 7.5 (cath BA7.5) were determined by a fluorogenic assay using synthetic substrate Z-Arg-Arg-AMC. The immunostaining pattern of cath B was determined in 239 lung tumour tissue sections, showing the presence of the enzyme in tumour cells (cath BT-I) and in tumour-associated histiocytes (cath BH-I). The median levels of cath BAT, cath BA7.5 and cath BC were 5.6-, 3.2- and 9.1-fold higher (P < 0.001), respectively, in tumour tissue than in non-tumourous lung parenchyma. Out of 131 tissue sections from patients with squamous cell carcinoma (SCC), 59.5% immunostained positively for cath B, while among the 108 adenocarcinoma (AC) patients 48.2% of tumours showed a positive reaction. There was a strong relationship between the levels of cath BAT, cath BA7.5, cath BC and cath BT-I in the primary tumours and the presence of lymph node metastases. Significant correlation with overall survival was observed for cath BT-I and cath BA7.5 (P < 0.01 and P < 0.05, respectively) in patients suffering from SCC. In these patients positive cath B in tumour cells (cath BT-I) and negative cath B in histiocytes (cath BH-I) indicated significantly shorter survival rate compared with patients with negative cath BT-I and positive cath BH-I (P < 0.0001). In contrast, in AC patients, both, positive cath BT-I and positive cath BH-I, indicated poor survival probability (P < 0.014). From these results we conclude that the proteolytic enzyme cath B is an independent prognostic factor for overall survival of patients suffering from SCC of the lung. © 1999 Cancer Research Campaign

Keywords: cathepsin B, prognosis, non-small cell lung cancer, squamous cell carcinoma

Full Text

The Full Text of this article is available as a PDF (269K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Abel U, Berger J, Wiebelt H. CRITLEVEL: an exploratory procedure for the evaluation of quantitative prognostic factors. Methods Inf Med. 1984 Jul;23(3):154–156. [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Budihna M, Strojan P, Smid L, Skrk J, Vrhovec I, Zupevc A, Rudolf Z, Zargi M, Krasovec M, Svetic B, et al. Prognostic value of cathepsins B, H, L, D and their endogenous inhibitors stefins A and B in head and neck carcinoma. Biol Chem Hoppe Seyler. 1996 Jun;377(6):385–390. [PubMed]
  • Calkins CC, Sloane BF. Mammalian cysteine protease inhibitors: biochemical properties and possible roles in tumor progression. Biol Chem Hoppe Seyler. 1995 Feb;376(2):71–80. [PubMed]
  • Castiglioni T, Merino MJ, Elsner B, Lah TT, Sloane BF, Emmert-Buck MR. Immunohistochemical analysis of cathepsins D, B, and L in human breast cancer. Hum Pathol. 1994 Sep;25(9):857–862. [PubMed]
  • Ebert W, Knoch H, Werle B, Trefz G, Muley T, Spiess E. Prognostic value of increased lung tumor tissue cathepsin B. Anticancer Res. 1994 May-Jun;14(3A):895–899. [PubMed]
  • Emmert-Buck MR, Roth MJ, Zhuang Z, Campo E, Rozhin J, Sloane BF, Liotta LA, Stetler-Stevenson WG. Increased gelatinase A (MMP-2) and cathepsin B activity in invasive tumor regions of human colon cancer samples. Am J Pathol. 1994 Dec;145(6):1285–1290. [PubMed]
  • Erdel M, Trefz G, Spiess E, Habermaas S, Spring H, Lah T, Ebert W. Localization of cathepsin B in two human lung cancer cell lines. J Histochem Cytochem. 1990 Sep;38(9):1313–1321. [PubMed]
  • Fidler IJ. Inhibition of pulmonary metastasis by intravenous injection of specifically activated macrophages. Cancer Res. 1974 May;34(5):1074–1078. [PubMed]
  • Higashiyama M, Doi O, Kodama K, Yokouchi H, Tateishi R. Cathepsin B expression in tumour cells and laminin distribution in pulmonary adenocarcinoma. J Clin Pathol. 1993 Jan;46(1):18–22. [PMC free article] [PubMed]
  • Honn KV, Timár J, Rozhin J, Bazaz R, Sameni M, Ziegler G, Sloane BF. A lipoxygenase metabolite, 12-(S)-HETE, stimulates protein kinase C-mediated release of cathepsin B from malignant cells. Exp Cell Res. 1994 Sep;214(1):120–130. [PubMed]
  • Hsu SM, Raine L, Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem. 1981 Apr;29(4):577–580. [PubMed]
  • Hughes SJ, Glover TW, Zhu XX, Kuick R, Thoraval D, Orringer MB, Beer DG, Hanash S. A novel amplicon at 8p22-23 results in overexpression of cathepsin B in esophageal adenocarcinoma. Proc Natl Acad Sci U S A. 1998 Oct 13;95(21):12410–12415. [PubMed]
  • Inoue T, Ishida T, Sugio K, Sugimachi K. Cathepsin B expression and laminin degradation as factors influencing prognosis of surgically treated patients with lung adenocarcinoma. Cancer Res. 1994 Dec 1;54(23):6133–6136. [PubMed]
  • Kos J, Smid A, Krasovec M, Svetic B, Lenarcic B, Vrhovec I, Skrk J, Turk V. Lysosomal proteases cathepsins D, B, H, L and their inhibitors stefins A and B in head and neck cancer. Biol Chem Hoppe Seyler. 1995 Jul;376(7):401–405. [PubMed]
  • Kos J, Stabuc B, Schweiger A, Krasovec M, Cimerman N, Kopitar-Jerala N, Vrhovec I. Cathepsins B, H, and L and their inhibitors stefin A and cystatin C in sera of melanoma patients. Clin Cancer Res. 1997 Oct;3(10):1815–1822. [PubMed]
  • Kos J, Nielsen HJ, Krasovec M, Christensen IJ, Cimerman N, Stephens RW, Brünner N. Prognostic values of cathepsin B and carcinoembryonic antigen in sera of patients with colorectal cancer. Clin Cancer Res. 1998 Jun;4(6):1511–1516. [PubMed]
  • Krepela E, Kasafírek E, Novák K, Viklický J. Increased cathepsin B activity in human lung tumors. Neoplasma. 1990;37(1):61–70. [PubMed]
  • Lah TT, Kos J. Cysteine proteinases in cancer progression and their clinical relevance for prognosis. Biol Chem. 1998 Feb;379(2):125–130. [PubMed]
  • Lah TT, Kokalj-Kunovar M, Strukelj B, Pungercar J, Barlic-Maganja D, Drobnic-Kosorok M, Kastelic L, Babnik J, Golouh R, Turk V. Stefins and lysosomal cathepsins B, L and D in human breast carcinoma. Int J Cancer. 1992 Jan 2;50(1):36–44. [PubMed]
  • Lah TT, Calaf G, Kalman E, Shinde BG, Russo J, Jarosz D, Zabrecky J, Somers R, Daskal I. Cathepsins D, B and L in breast carcinoma and in transformed human breast epithelial cells (HBEC). Biol Chem Hoppe Seyler. 1995 Jun;376(6):357–363. [PubMed]
  • Lah TT, Hawley M, Rock KL, Goldberg AL. Gamma-interferon causes a selective induction of the lysosomal proteases, cathepsins B and L, in macrophages. FEBS Lett. 1995 Apr 17;363(1-2):85–89. [PubMed]
  • Ledakis P, Tester WT, Rosenberg N, Romero-Fischmann D, Daskal I, Lah TT. Cathepsins D, B, and L in malignant human lung tissue. Clin Cancer Res. 1996 Mar;2(3):561–568. [PubMed]
  • Leto G, Tumminello FM, Pizzolanti G, Montalto G, Soresi M, Carroccio A, Ippolito S, Gebbia N. Lysosomal aspartic and cysteine proteinases serum levels in patients with pancreatic cancer or pancreatitis. Pancreas. 1997 Jan;14(1):22–27. [PubMed]
  • Mori M, Kohli A, Baker SP, Savas L, Fraire AE. Laminin and cathepsin B as prognostic factors in stage I non-small cell lung cancer: are they useful? Mod Pathol. 1997 Jun;10(6):572–577. [PubMed]
  • Mantovani A. Tumor-associated macrophages. Curr Opin Immunol. 1989;2(5):689–692. [PubMed]
  • Mikkelsen T, Yan PS, Ho KL, Sameni M, Sloane BF, Rosenblum ML. Immunolocalization of cathepsin B in human glioma: implications for tumor invasion and angiogenesis. J Neurosurg. 1995 Aug;83(2):285–290. [PubMed]
  • Opdenakker G, Van Damme J. Chemotactic factors, passive invasion and metastasis of cancer cells. Immunol Today. 1992 Nov;13(11):463–464. [PubMed]
  • Ozeki Y, Takishima K, Takagi K, Aida S, Tamai S, Mamiya G, Ogata T. Immunohistochemical analysis of cathepsin B expression in human lung adenocarcinoma: the role in cancer progression. Jpn J Cancer Res. 1993 Sep;84(9):972–975. [PubMed]
  • Rempel SA, Rosenblum ML, Mikkelsen T, Yan PS, Ellis KD, Golembieski WA, Sameni M, Rozhin J, Ziegler G, Sloane BF. Cathepsin B expression and localization in glioma progression and invasion. Cancer Res. 1994 Dec 1;54(23):6027–6031. [PubMed]
  • Rozhin J, Robinson D, Stevens MA, Lah TT, Honn KV, Ryan RE, Sloane BF. Properties of a plasma membrane-associated cathepsin B-like cysteine proteinase in metastatic B16 melanoma variants. Cancer Res. 1987 Dec 15;47(24 Pt 1):6620–6628. [PubMed]
  • Rozhin J, Sameni M, Ziegler G, Sloane BF. Pericellular pH affects distribution and secretion of cathepsin B in malignant cells. Cancer Res. 1994 Dec 15;54(24):6517–6525. [PubMed]
  • Sedo A, Krepela E, Kasafírek E. Dipeptidyl peptidase IV, prolyl endopeptidase and cathepsin B activities in primary human lung tumors and lung parenchyma. J Cancer Res Clin Oncol. 1991;117(3):249–253. [PubMed]
  • Shuja S, Sheahan K, Murnane MJ. Cysteine endopeptidase activity levels in normal human tissues, colorectal adenomas and carcinomas. Int J Cancer. 1991 Sep 30;49(3):341–346. [PubMed]
  • Sloane BF, Rozhin J, Hatfield JS, Crissman JD, Honn KV. Plasma membrane-associated cysteine proteinases in human and animal tumors. Exp Cell Biol. 1987;55(4):209–224. [PubMed]
  • Sloane BF, Moin K, Sameni M, Tait LR, Rozhin J, Ziegler G. Membrane association of cathepsin B can be induced by transfection of human breast epithelial cells with c-Ha-ras oncogene. J Cell Sci. 1994 Feb;107(Pt 2):373–384. [PubMed]
  • Stonelake PS, Jones CE, Neoptolemos JP, Baker PR. Proteinase inhibitors reduce basement membrane degradation by human breast cancer cell lines. Br J Cancer. 1997;75(7):951–959. [PMC free article] [PubMed]
  • Strohmaier AR, Porwol T, Acker H, Spiess E. Tomography of cells by confocal laser scanning microscopy and computer-assisted three-dimensional image reconstruction: localization of cathepsin B in tumor cells penetrating collagen gels in vitro. J Histochem Cytochem. 1997 Jul;45(7):975–983. [PubMed]
  • Sukoh N, Abe S, Ogura S, Isobe H, Takekawa H, Inoue K, Kawakami Y. Immunohistochemical study of cathepsin B. Prognostic significance in human lung cancer. Cancer. 1994 Jul 1;74(1):46–51. [PubMed]
  • Sukoh N, Abe S, Nakajima I, Ogura S, Isobe H, Inoue K, Kawakami Y. Immunohistochemical distributions of cathepsin B and basement membrane antigens in human lung adenocarcinoma: association with invasion and metastasis. Virchows Arch. 1994;424(1):33–38. [PubMed]
  • Bandlow G, Härtl R. Selection and isolation of a new variant of DBA/2 mastocytoma P 815 X 2. J Cancer Res Clin Oncol. 1989;115(6):550–553. [PubMed]
  • Ulbricht B, Hagmann W, Ebert W, Spiess E. Differential secretion of cathepsins B and L from normal and tumor human lung cells stimulated by 12(S)-hydroxy-eicosatetraenoic acid. Exp Cell Res. 1996 Aug 1;226(2):255–263. [PubMed]
  • Werle B, Ebert W, Klein W, Spiess E. Cathepsin B in tumors, normal tissue and isolated cells from the human lung. Anticancer Res. 1994 May-Jun;14(3A):1169–1176. [PubMed]
  • Werle B, Ebert W, Klein W, Spiess E. Charge polymorphism in human lung cell pro-cathepsin B. Anticancer Res. 1996 Jan-Feb;16(1):49–53. [PubMed]
  • Werle B, Jülke B, Lah T, Spiess E, Ebert W. Cathepsin B fraction active at physiological pH of 7.5 is of prognostic significance in squamous cell carcinoma of human lung. Br J Cancer. 1997;75(8):1137–1143. [PMC free article] [PubMed]
  • Yan S, Sameni M, Sloane BF. Cathepsin B and human tumor progression. Biol Chem. 1998 Feb;379(2):113–123. [PubMed]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK