Abstract
Cystatin E/M (CST6) is a natural inhibitor of lysosomal cysteine proteases. Recent studies have shown that experimental manipulation of CST6 expression alters the metastatic behavior of human breast cancer cells. However, the association of CST6 with prostate cancer invasion and progression remains unclear. Here, we show that CST6 is robustly expressed in normal human prostate epithelium, whereas its expression is downregulated in metastatic prostate cell lines and prostate tumor tissues. Treatment of metastatic prostate cell lines with the histone deacetylase inhibitor trichostatin A resulted in significant induction of CST6 mRNA levels and increased CST6 protein expression, indicating that epigenetic silencing may play a role in the loss of CST6 expression observed in prostate cancer. CST6 overexpression in human prostate cancer cells significantly reduced in vitro cell proliferation and matrigel invasion. Furthermore, the results from a bioluminescence tumor/metastasis model showed that the overexpression of CST6 significantly inhibits tumor growth and the incidence of lung metastasis. These results suggest that the downregulation of the CST6 gene is associated with promoter histone modifications and that this association plays an important role in prostate cancer progression during the invasive and metastatic stages of the disease.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Abrahamson M . (1994). Cystatins. Methods Enzymol 244: 685–700.
Ai L, Kim WJ, Kim TY, Fields CR, Massoll NA, Robertson KD et al. (2006). Epigenetic silencing of the tumor suppressor cystatin M occurs during breast cancer progression. Cancer Res 66: 7899–7909.
Alvarez-Fernandez M, Barrett AJ, Gerhartz B, Dando PM, Ni J, Abrahamson M . (1999). Inhibition of mammalian legumain by some cystatins is due to a novel second reactive site. J Biol Chem 274: 19195–19203.
Bogenrieder T, Herlyn M . (2003). Axis of evil: molecular mechanisms of cancer metastasis. Oncogene 22: 6524–6536.
Brunner N, Pyke C, Hansen CH, Romer J, Grondahl-Hansen J, Dano K . (1994). Urokinase plasminogen activator (uPA) and its type 1 inhibitor (PAI-1): regulators of proteolysis during cancer invasion and prognostic parameters in breast cancer. Cancer Treat Res 71: 299–309.
Cameron EE, Bachman KE, Myohanen S, Herman JG, Baylin SB . (1999). Synergy of demethylation and histone deacetylase inhibition in the re-expression of genes silenced in cancer. Nat Genet 21: 103–107.
DeClerck YA, Imren S . (1994). Protease inhibitors: role and potential therapeutic use in human cancer. Eur J Cancer 30A: 2170–2180.
Esteller M, Corn PG, Baylin SB, Herman JG . (2001). A gene hypermethylation profile of human cancer. Cancer Res 61: 3225–3229.
Hake SB, Xiao A, Allis CD . (2004). Linking the epigenetic ‘language’ of covalent histone modifications to cancer. Br J Cancer 90: 761–769.
Keppler D, Sloane BF . (1996). Cathepsin B: multiple enzyme forms from a single gene and their relation to cancer. Enzyme Protein 49: 94–105.
Kim TY, Zhong S, Fields CR, Kim JH, Robertson KD . (2006). Epigenomic profiling reveals novel and frequent targets of aberrant DNA methylation-mediated silencing in malignant glioma. Cancer Res 66: 7490–7501.
Konduri SD, Yanamandra N, Siddique K, Joseph A, Dinh DH, Olivero WC et al. (2002). Modulation of cystatin C expression impairs the invasive and tumorigenic potential of human glioblastoma cells. Oncogene 21: 8705–8712.
Krueger S, Kellner U, Buehling F, Roessner A . (2001). Cathepsin L antisense oligonucleotides in a human osteosarcoma cell line: effects on the invasive phenotype. Cancer Gene Ther 8: 522–528.
Marks P, Rifkind RA, Richon VM, Breslow R, Miller T, Kelly WK . (2001a). Histone deacetylases and cancer: causes and therapies. Nat Rev Cancer 1: 194–202.
Marks PA, Rifkind RA, Richon VM, Breslow R . (2001b). Inhibitors of histone deacetylase are potentially effective anticancer agents. Clin Cancer Res 7: 759–760.
Ni J, Abrahamson M, Zhang M, Fernandez MA, Grubb A, Su J et al. (1997). Cystatin E is a novel human cysteine proteinase inhibitor with structural resemblance to family 2 cystatins. J Biol Chem 272: 10853–10858.
Pulukuri SM, Gorantla B, Rao JS . (2007a). Inhibition of histone deacetylase activity promotes invasion of human cancer cells through activation of urokinase plasminogen activator (uPA). J Biol Chem 282: 35594–35603.
Pulukuri SM, Patibandla S, Patel J, Estes N, Rao JS . (2007b). Epigenetic inactivation of the tissue inhibitor of metalloproteinase-2 (TIMP-2) gene in human prostate tumors. Oncogene 26: 5229–5237.
Pulukuri SM, Rao JS . (2007). Small interfering RNA directed reversal of urokinase plasminogen activator demethylation inhibits prostate tumor growth and metastasis. Cancer Res 67: 6637–6646.
Qiu J, Ai L, Ramachandran C, Yao B, Gopalakrishnan S, Fields CR et al. (2008). Invasion suppressor cystatin E/M (CST6): high-level cell type-specific expression in normal brain and epigenetic silencing in gliomas. Lab Invest 88: 910–925.
Richon VM, Sandhoff TW, Rifkind RA, Marks PA . (2000). Histone deacetylase inhibitor selectively induces p21WAF1 expression and gene-associated histone acetylation. Proc Natl Acad Sci USA 97: 10014–10019.
Roshy S, Sloane BF, Moin K . (2003). Pericellular cathepsin B and malignant progression. Cancer Metastasis Rev 22: 271–286.
Sambucetti LC, Fischer DD, Zabludoff S, Kwon PO, Chamberlin H, Trogani N et al. (1999). Histone deacetylase inhibition selectively alters the activity and expression of cell cycle proteins leading to specific chromatin acetylation and antiproliferative effects. J Biol Chem 274: 34940–34947.
Schagdarsurengin U, Pfeifer GP, Dammann R . (2007). Frequent epigenetic inactivation of cystatin M in breast carcinoma. Oncogene 26: 3089–3094.
Shah RB, Mehra R, Chinnaiyan AM, Shen R, Ghosh D, Zhou M et al. (2004). Androgen-independent prostate cancer is a heterogeneous group of diseases: lessons from a rapid autopsy program. Cancer Res 64: 9209–9216.
Shridhar R, Zhang J, Song J, Booth BA, Kevil CG, Sotiropoulou G et al. (2004). Cystatin M suppresses the malignant phenotype of human MDA-MB-435S cells. Oncogene 23: 2206–2215.
Sotiropoulou G, Anisowicz A, Sager R . (1997). Identification, cloning, and characterization of cystatin M, a novel cysteine proteinase inhibitor, down-regulated in breast cancer. J Biol Chem 272: 903–910.
Turk B, Turk D, Turk V . (2000). Lysosomal cysteine proteases: more than scavengers. Biochim Biophys Acta 1477: 98–111.
Turk V, Bode W . (1991). The cystatins: protein inhibitors of cysteine proteinases. FEBS Lett 285: 213–219.
Veena MS, Lee G, Keppler D, Mendonca MS, Redpath JL, Stanbridge EJ et al. (2008). Inactivation of the cystatin E/M tumor suppressor gene in cervical cancer. Genes Chromosomes Cancer 47: 740–754.
Yanamandra N, Gumidyala KV, Waldron KG, Gujrati M, Olivero WC, Dinh DH et al. (2004). Blockade of cathepsin B expression in human glioblastoma cells is associated with suppression of angiogenesis. Oncogene 23: 2224–2230.
Yoon SO, Kim MM, Chung AS . (2001). Inhibitory effect of selenite on invasion of HT1080 tumor cells. J Biol Chem 276: 20085–20092.
Zhang J, Shridhar R, Dai Q, Song J, Barlow SC, Yin L et al. (2004). Cystatin M: a novel candidate tumor suppressor gene for breast cancer. Cancer Res 64: 6957–6964.
Zhong S, Fields CR, Su N, Pan YX, Robertson KD . (2007). Pharmacologic inhibition of epigenetic modifications, coupled with gene expression profiling, reveals novel targets of aberrant DNA methylation and histone deacetylation in lung cancer. Oncogene 26: 2621–2634.
Acknowledgements
We are grateful to Dr Hnilica of the Department of Pathology at the University of Illinois College of Medicine (Peoria) for kindly providing normal and tumor human prostate tissues. We thank Shellee Abraham for preparing the paper and Diana Meister and Sushma Jasti for paper review. We also thank Noorjehan Ali and Lavanya Talluri, for technical assistance. This research was supported by National Cancer Institute Grant CA75557, CA92393, CA95058, CA116708, CA138409, N.I.N.D.S. NS47699, NS57529 and NS61835 and Caterpillar Inc., OSF St Francis Inc. Peoria, IL (to JSR). The contents are solely the responsibility of the authors and do not necessarily represent the official views of NIH.
Author information
Authors and Affiliations
Corresponding author
Additional information
Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)
Rights and permissions
About this article
Cite this article
Pulukuri, S., Gorantla, B., Knost, J. et al. Frequent loss of cystatin E/M expression implicated in the progression of prostate cancer. Oncogene 28, 2829–2838 (2009). https://doi.org/10.1038/onc.2009.134
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2009.134