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Selective suppression of cathepsin L by antisense cDNA impairs human brain tumor cell invasion in vitro and promotes apoptosis

Cancer Gene Therapy volume 10, pages 141–151 (2003)Cite this article

Abstract

Invasion and metastasis of certain tumors are accompanied by increased mRNA protein levels and enzymatic activity of cathepsin L. Cathepsin L has also been suggested to play a role in the proteolytic cascades associated with apoptosis. To investigate the role of cathepsin L in brain tumor invasion and apoptosis, the human glioma cell line, IPTP, was stably transfected with full-length antisense and sense cDNA of cathepsin L. Down-regulation of cathepsin L by antisense cDNA significantly impaired (up to 70%) glioma cell invasion in vitro and markedly increased glioma cell apoptosis induced by staurosporine. Compared to control and parental cell lines, antisense down-regulation of cathepsin L was associated with an earlier induction of caspase-3 activity. Up-regulation of cathepsin L activity by sense cDNA was associated with reduced apoptosis and later induction of caspase-3 activity. Moreover, down-regulation of cathepsin L lowered the expression of antiapoptotic protein Bcl-2, whereas up-regulation increased the expression of Bcl-2, indicating that cathepsin L acts upstream of caspase-3. These data show that cathepsin L is an important protein mediating the malignancy of gliomas and its inhibition may diminish their invasion and lead to increased tumor cell apoptosis by reducing apoptotic threshold.

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Acknowledgements

This study was supported by the Ministry of Education, Science and Sport of Republic of Slovenia (program no. 0105-509, to TL); the PhD fellowship to N L by the Ministry of Education, Science and Sport; the British Council grant support for Partnerships in Science PSP 6; and the Samantha Dickinson Research Trust (G J P) and the European Union (G J P). We thank Astrid Fitter (Department of Pediatric Hematology and Oncology, Hannover Medical School) for her excellent technical assistance.

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Authors and Affiliations

  1. Department of Genetic Toxicology and Cancer Biology, National Institute of Biology, Ljubljana, Slovenia

    Nataša Levičar & Tamara T Lah

  2. Department of Neuropathology, Experimental Neuro-Oncology Group, Institute of Psychiatry, King's College London, De Crespigny Park, Denmark Hill, London, UK

    Nataša Levičar, Emma Daley & Geoffrey J Pilkington

  3. Department of Pediatric Hematology and Oncology, Hannover Medical School, Hannover, Germany

    Ricardo A Dewey

  4. MRC Applied Neuroscience Group, School of Biomedical Sciences, University of Nottingham, Queen's Medical Centre, Nottingham, UK

    Timothy E Bates

  5. FACS Laboratory, London Research Institute, Lincoln's Inn Fields Laboratories, Cancer Research, 44 Lincoln's Inn Fields, London, UK

    Derek Davies

  6. Department of Biochemical Research and Drug Design, Krka, d.d., Cesta na Brdo 49, Ljubljana, Slovenia

    Janko Kos

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  1. Nataša Levičar
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Correspondence to Tamara T Lah.

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Levičar, N., Dewey, R., Daley, E. et al. Selective suppression of cathepsin L by antisense cDNA impairs human brain tumor cell invasion in vitro and promotes apoptosis. Cancer Gene Ther 10, 141–151 (2003). https://doi.org/10.1038/sj.cgt.7700546

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