Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Inhibition of cathepsin B and MMP-9 gene expression in glioblastoma cell line via RNA interference reduces tumor cell invasion, tumor growth and angiogenesis

Abstract

Extracellular proteases have been shown to cooperatively influence matrix degradation and tumor cell invasion through proteolytic cascades, with individual proteases having distinct roles in tumor growth, invasion, migration and angiogenesis. Matrix metalloproteases (MMP)-9 and cathepsin B have been shown to participate in the processes of tumor growth, vascularization and invasion of gliomas. In the present study, we used a cytomegalovirus promoter-driven DNA template approach to induce hairpin RNA (hpRNA)-triggered RNA interference (RNAi) to block MMP-9 and cathepsin B gene expression with a single construct. Transfection of a plasmid vector-expressing double-stranded RNA (dsRNA) for MMP-9 and cathepsin B significantly inhibited MMP-9 and cathepsin B expression and reduced the invasive behavior of SNB19, glioblastoma cell line in Matrigel and spheroid invasion models. Downregulation of MMP-9 and cathepsin B using RNAi in SNB19 cells reduced cell–cell interaction of human microvascular endothelial cells, resulting in the disruption of capillary network formation in both in vitro and in vivo models. Direct intratumoral injections of plasmid DNA expressing hpRNA for MMP-9 and cathepsin B significantly inhibited established glioma tumor growth and invasion in intracranial tumors in vivo. Further intraperitoneal (ip) injections of plasmid DNA expressing hpRNA for MMP-9 and cathepsin B completely regressed pre-established tumors for a long time (4 months) without any indication of these tumor cells. For the first time, these observations demonstrate that the simultaneous RNAi-mediated targeting of MMP-9 and cathepsin B has potential application for the treatment of human gliomas.

This is a preview of subscription content, access via your institution

Access options

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

Scheme 1
Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

Abbreviations

MMP-9:

matrix metalloproteases -9

RNAi:

RNA interference vector expressing siRNA for cathepsin B and MMP-9 (pCM)

CMV:

cytomegalovirus

SV40:

simian virus type 40

PCR:

polymerase chain reaction

PBS:

phosphate-buffered saline

FITC:

fluoresceine-5-isothiocyanate

DiI:

1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanineperchlorate

DiO:

3,3′-dioctadecyloxacarbocyanine perchlorate

GFP:

green fluorescent protein

ECM:

extracellular matrix

References

  • Arkona C and Wiederanders B . (1996). Biol. Chem., 377, 695–702.

  • Bergers G, Brekken R, McMahon G, Vu TH, Itoh T, Tamaki K, Tanzawa K, Thorpe P, Itohara S, Werb Z and Hanahan D . (2000). Nat. Cell Biol., 2, 737–744.

  • Bertrand JR, Pottier M, Vekris A, Opolon P, Maksimenko A and Malvy C . (2002). Biochem. Biophys. Res. Commun., 296, 1000–1004.

  • Bervar A, Zajc I, Sever N, Katunuma N, Sloane BF and Lah TT . (2003). Biol. Chem., 384, 447–455.

  • Boike G, Lah T, Sloane BF, Rozhin J, Honn K, Guirguis R, Stracke ML, Liotta LA and Schiffmann E . (1992). Melanoma Res., 1, 333–340.

  • Chandrasekar N, Jasti S, Alfred-Yung WK, Ali-Osman F, Dinh DH, Olivero WC, Gujrati M, Kyritsis AP, Nicolson GL, Rao JS and Mohanam S . (2000). Clin. Exp. Metast., 18, 337–342.

  • Coussens LM, Tinkle CL, Hanahan D and Werb Z . (2000). Cell, 103, 481–490.

  • Egeblad M and Werb Z . (2002). Nat. Rev. Cancer, 2, 161–174.

  • Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K and Tuschl T . (2001). Nature, 411, 494–498.

  • Emmert-Buck MR, Roth MJ, Zhuang Z, Campo E, Rozhin J, Sloane BF, Liotta LA and Stetler-Stevenson WG . (1994). Am. J. Pathol., 145, 1285–1290.

  • Go Y, Chintala SK, Mohanam S, Gokaslan Z, Venkaiah B, Bjerkvig R, Oka K, Nicolson GL, Sawaya R and Rao JS . (1997). Clin. Exp. Metast., 15, 440–446.

  • Guinec N, Dalet-Fumeron V and Pagano M . (1993). Biol. Chem. Hoppe Seyler, 374, 1135–1146.

  • Hammond SM, Boettcher S, Caudy AA, Kobayashi R and Hannon GJ . (2001). Science, 293, 1146–1150.

  • Himelstein BP, Canete-Soler R, Bernhard EJ, Dilks DW and Muschel RJ . (1994). Invas. Metast., 14, 246–258.

  • Hua J and Muschel RJ . (1996). Cancer Res., 56, 5279–5284.

  • Itoh T, Tanioka M, Matsuda H, Nishimoto H, Yoshioka T, Suzuki R and Uehira M . (1999). Clin. Exp. Metast., 17, 177–181.

  • Jen KY and Gewirtz AM . (2000). Stem Cells, 18, 307–319.

  • Kachra Z, Beaulieu E, Delbecchi L, Mousseau N, Berthelet F, Moumdjian R, Del Maestro R and Beliveau R . (1999). Clin. Exp. Metast., 17, 555–566.

  • Kobayashi H, Moniwa N, Sugimura M, Shinohara H, Ohi H and Terao T . (1993). Biochim. Biophys. Acta, 1178, 55–62.

  • Kobayashi H, Ohi H, Sugimura M, Shinohara H, Fujii T and Terao T . (1992). Cancer Res., 52, 3610–3614.

  • Kondraganti S, Mohanam S, Chintala SK, Kin Y, Jasti SL, Nirmala C, Lakka SS, Adachi Y, Kyritsis AP, Ali-Osman F, Sawaya R, Fuller GN and Rao JS . (2000). Cancer Res., 60, 6851–6855.

  • Konduri SD, Rao CN, Chandrasekar N, Tasiou A, Mohanam S, Kin Y, Lakka SS, Dinh D, Olivero WC, Gujrati M, Foster DC, Kisiel W and Rao JS . (2001). Oncogene, 20, 6938–6945.

  • Lakka SS, Jasti SL, Kyritsis AP, Yung WK, Ali-Osman F, Nicolson GL and Rao JS . (2000). Clin. Exp. Metast., 18, 245–252.

  • Lakka SS, Rajan M, Gondi C, Yanamandra N, Chandrasekar N, Jasti SL, Adachi Y, Siddique K, Gujrati M, Olivero W, Dinh DH, Kouraklis G, Kyritsis AP and Rao JS . (2002). Oncogene, 21, 8011–8019.

  • Leunig M, Yuan F, Menger MD, Boucher Y, Goetz AE, Messmer K and Jain RK . (1992). Cancer Res., 52, 6553–6560.

  • Lewis DL, Hagstrom JE, Loomis AG, Wolff JA and Herweijer H . (2002). Nat. Genet., 32, 107–108.

  • Liotta LA, Rao CN and Wewer UM . (1986). Annu. Rev. Biochem., 55, 1037–1057.

  • Mai J, Sameni M, Mikkelsen T and Sloane BF . (2002). Biol. Chem., 383, 1407–1413.

  • McCaffrey AP, Meuse L, Pham TT, Conklin DS, Hannon GJ and Kay MA . (2002). Nature, 418, 38–39.

  • Miyagishi M, Hayashi M and Taira K . (2003). Antisense Nucleic Acid Drug Dev., 13, 1–7.

  • Mohanam S, Jasti SL, Kondraganti SR, Chandrasekar N, Lakka SS, Kin Y, Fuller GN, Yung AW, Kyritsis AP, Dinh DH, Olivero WC, Gujrati M, Ali-Osman F and Rao JS . (2001). Oncogene, 20, 3665–3673.

  • Pozzi A, LeVine WF and Gardner HA . (2002). Oncogene, 21, 272–281.

  • Premzl A, Zavasnik-Bergant V, Turk V and Kos J . (2003). Exp. Cell Res., 283, 206–214.

  • Raithatha SA, Muzik H, Muzik H, Rewcastle NB, Johnston RN, Edwards DR and Forsyth PA . (2000). Neuro-oncology, 2, 145–150.

  • Rao JS, Steck PA, Mohanam S, Stetler-Stevenson WG, Liotta LA and Sawaya R . (1993). Cancer Res., 53, 2208–2211.

  • Redwood SM, Liu BC, Weiss RE, Hodge DE and Droller MJ . (1992). Cancer, 69, 1212–1219.

  • Rooprai HK, Van Meter T, Rucklidge GJ, Hudson L, Everall IP and Pilkington GJ . (1998). Int. J. Oncol., 13, 1153–1157.

  • Simon C, Goepfert H and Boyd D . (1998). Cancer Res., 58, 1135–1139.

  • Sloane BF, Rozhin J, Johnson K, Taylor H, Crissman JD and Honn KV . (1986). Proc. Natl. Acad. Sci. USA, 83, 2483–2487.

  • Stark GR, Kerr IM, Williams BR, Silverman RH and Schreiber RD . (1998). Annu. Rev. Biochem., 67, 227–264.

  • Vu TH, Shipley JM, Bergers G, Berger JE, Helms JA, Hanahan D, Shapiro SD, Senior RM and Werb Z . (1998). Cell, 93, 411–422.

  • Yu Q and Stamenkovic I . (1999). Genes Dev., 13, 35–48.

  • Zamore PD . (2001). Nat. Struct. Biol., 8, 746–750.

Download references

Acknowledgements

We thank Karen Minter for preparing the manuscript, and Sushma Jasti and Diana Meister for manuscript review. This research was supported by the National Cancer Institute CA 85216 and CA 75557 and the National Institute of Neurological Disorders and Stroke Grant NS47699 (to JSR).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jasti S Rao.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lakka, S., Gondi, C., Yanamandra, N. et al. Inhibition of cathepsin B and MMP-9 gene expression in glioblastoma cell line via RNA interference reduces tumor cell invasion, tumor growth and angiogenesis. Oncogene 23, 4681–4689 (2004). https://doi.org/10.1038/sj.onc.1207616

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1207616

Keywords

This article is cited by

Search

Quick links