This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Szpaderska, A. M. Articles by Frankfater, A. Search for Related Content PubMed PubMed Citation Articles by Szpaderska, A. M. Articles by Frankfater, A.

An Intracellular Form of Cathepsin B Contributes to Invasiveness in Cancer

Division of Molecular and Cellular Biochemistry, Department of Cell Biology, Neurobiology and Anatomy, Loyola University Medical Center, Maywood, Illinois 60153

Cathepsin B is a lysosomal cysteine proteinase whose expression and trafficking are frequently altered in cancer, and plasma membrane and secreted forms are thought to contribute to the invasive and metastatic properties of malignant tumors. We have manipulated the expression of cathepsin B in several tumor cell lines and measured their capacity to invade through a reconstituted extracellular (Matrigel) matrix. Transient expression of human cathepsin B in a poorly metastatic B16F1 murine melanoma variant produced a 3–5-fold increase in cathepsin B activity and a comparable increase in invasiveness. Stable antisense cathepsin B-expressing clones of the highly metastatic human melanoma A375M and prostate carcinoma PC3M cell lines produced 40–50% less cathepsin B than control cells and were proportionately less invasive. In contrast, manipulating cathepsin B levels had no effect on cell migration across an uncoated membrane. The anionic cathepsin B inhibitor (L-3-trans-propylcarbamoyloxirane-2-carbony)-L-isoleucyl-L-proline (CA-074), at a concentration of 1 µM, caused a nearly quantitative inhibition of extracellular cathepsin B but had no effect on Matrigel invasion. In contrast, the equally potent but less selective inhibitor, trans-epoxysuccinyl-L-leucylamino(4-guanidino)butane (E-64) inhibited invasion by 75%. Surprisingly, at a concentration of 10 µM, CA-074 slowly permeated the cells, causing an 80–95% inhibition of intracellular cathepsin B after 12 h, the duration of the invasion assay. The membrane-permeant cathepsin B inhibitor, CA-074 methyl ester, and the higher concentration of CA-074 that inhibited intracellular cathepsin B both significantly reduced Matrigel invasion. Collectively, these results identify an intracellular role for cathepsin B in matrix degradation. They also indicate that caution should be exercised in assuming that CA-074 is unable to enter cells when it is used to inhibit biological processes of long duration.

This article has been cited by other articles:

				S.-D. Ha, A. Martins, K. Khazaie, J. Han, B. M. C. Chan, and S. O. Kim Cathepsin B Is Involved in the Trafficking of TNF-{alpha}-Containing Vesicles to the Plasma Membrane in Macrophages J. Immunol., July 1, 2008; 181(1): 690 - 697. [Abstract] [Full Text] [PDF]

				P. C. Ong, S. McGowan, M. C. Pearce, J. A. Irving, W.-T. Kan, S. A. Grigoryev, B. Turk, G. A. Silverman, K. Brix, S. P. Bottomley, et al. DNA Accelerates the Inhibition of Human Cathepsin V by Serpins J. Biol. Chem., December 21, 2007; 282(51): 36980 - 36986. [Abstract] [Full Text] [PDF]

				P. P. Rose, M. Bogyo, A. V. Moses, and K. Fruh Insulin-Like Growth Factor II Receptor-Mediated Intracellular Retention of Cathepsin B Is Essential for Transformation of Endothelial Cells by Kaposi's Sarcoma-Associated Herpesvirus J. Virol., August 1, 2007; 81(15): 8050 - 8062. [Abstract] [Full Text] [PDF]

				N. Muthukumaran, K. E. Miletti-Gonzalez, A. K. Ravindranath, and L. Rodriguez-Rodriguez Tumor Necrosis Factor-{alpha} Differentially Modulates CD44 Expression in Ovarian Cancer Cells Mol. Cancer Res., August 1, 2006; 4(8): 511 - 520. [Abstract] [Full Text] [PDF]

				O. Vasiljeva, A. Papazoglou, A. Kruger, H. Brodoefel, M. Korovin, J. Deussing, N. Augustin, B. S. Nielsen, K. Almholt, M. Bogyo, et al. Tumor cell-derived and macrophage-derived cathepsin B promotes progression and lung metastasis of mammary cancer. Cancer Res., May 15, 2006; 66(10): 5242 - 5250. [Abstract] [Full Text] [PDF]

				M. S. K. Sutherland, R. J. Sanderson, K. A. Gordon, J. Andreyka, C. G. Cerveny, C. Yu, T. S. Lewis, D. L. Meyer, R. F. Zabinski, S. O. Doronina, et al. Lysosomal Trafficking and Cysteine Protease Metabolism Confer Target-specific Cytotoxicity by Peptide-linked Anti-CD30-Auristatin Conjugates J. Biol. Chem., April 14, 2006; 281(15): 10540 - 10547. [Abstract] [Full Text] [PDF]

				W. Li, F. Ding, L. Zhang, Z. Liu, Y. Wu, A. Luo, M. Wu, M. Wang, Q. Zhan, and Z. Liu Overexpression of Stefin A in Human Esophageal Squamous Cell Carcinoma Cells Inhibits Tumor Cell Growth, Angiogenesis, Invasion, and Metastasis Clin. Cancer Res., December 15, 2005; 11(24): 8753 - 8762. [Abstract] [Full Text] [PDF]

				J. P. Sokol and W. P. Schiemann Cystatin C Antagonizes Transforming Growth Factor {beta} Signaling in Normal and Cancer Cells Mol. Cancer Res., March 1, 2004; 2(3): 183 - 195. [Abstract] [Full Text] [PDF]

				A.M. Szpaderska, J.D. Zuckerman, and L.A. DiPietro Differential Injury Responses in Oral Mucosal and Cutaneous Wounds Journal of Dental Research, August 1, 2003; 82(8): 621 - 626. [Abstract] [Full Text] [PDF]

				M. Mareel and A. Leroy Clinical, Cellular, and Molecular Aspects of Cancer Invasion Physiol Rev, April 1, 2003; 83(2): 337 - 376. [Abstract] [Full Text] [PDF]