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Inhibition of cathepsin L lowers the apoptotic threshold of glioblastoma cells by up-regulating p53 and transcription of caspases 3 and 7

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Abstract

Despite all the progress in cancer treatment, glioblastoma, the most malignant tumor of the central nervous system, remains a terminal disease and new therapeutic approaches are urgently needed. A combination of chemotherapy with modifications that lower the apoptotic threshold of cancer cells could be effective. Cathepsin L inhibition was suggested as one of such modifications but the mechanism of cathepsin L anti-apoptotic activity is largely unknown. In the present study we show that, in U87 glioblastoma cells, cathepsin L is present in the nucleus and regulates the transcription of effector caspases 3 and 7. In cells with low cathepsin L expression, p53 and prohibitin—transcription factors that regulate caspase 7 expression—accumulate in the nuclei. The importance of p53 in this process is highlighted by the fact that in U87 cells with inhibited p53 transcriptional activity or in p53-negative cells U251, cathepsin L inhibition did not influence caspase 7 expression and had minimal effect on the level of apoptosis. Since p53 pathways are often mutated in glioblastoma, the findings of our study need to be considered before using cathepsin L inhibition for glioblastoma therapy and suggest that such adjuvant therapy may be effective only for a subpopulation of p53 wild type glioblastoma patients.

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Acknowledgments

The work was supported by Slovenian Research Agency, Programme #P-0105-0245 (granted to TTL), and Young Researcher Project (granted to SK). The authors are thankful to Dr. Viktor Menart for human recombinant TNFα, Dr. Janko Kos for cathepsin L antibody and ELISA assay, Dr. Nobuhiko Katunuma for cathepsin L inhibitor Clik 148 and Dr. Bernd Wiederanders for the DNA-microarray.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

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

    Saša Kenig, Anja Pucer & Tamara Lah

  2. Institute of Physiology, Pharmacology and Toxicology, Veterinary Faculty, University of Ljubljana, Ljubljana, Slovenia

    Robert Frangež

  3. Department of Chemistry and Biochemistry, Faculty of Chemistry and Chemical Technology, University of Ljubljana, Ljubljana, Slovenia

    Tamara Lah

  4. ICGEB, Genome Stability, Padriciano 99, 34149, Trieste, Italy

    Saša Kenig

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  1. Saša Kenig
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Correspondence to Saša Kenig.

Electronic supplementary material

Below is the link to the electronic supplementary material.

10495_2011_600_MOESM1_ESM.tif

Electronic supplementary material 1: The efficacy of CatL silencing CatL expression was measured at the level of (a) mRNA and (b) enzyme activity in the control and transfected U87 cells. (a): U87 cells transfected with anti-CatL siRNA, non-si RNA and non-transfected control cells were collected with TRIzol reagent at designated times after transfection. CatL expression was determined by qRT-PCR. (b): Cells were collected in a homogenization buffer at designated times, proteins were extracted and enzyme activity tested using fluorescence-labeled substrate Z-Phe-Arg AMC, as decribed in Materials and methods. Results are presented as means of at least three independent experiments ± SD. Expression and activity in non-si cells is set as 1. (TIFF 25 kb)

Electronic supplementary material 2: The function of cathepsin L is not replaced by other proteases (TIFF 40 kb)

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Kenig, S., Frangež, R., Pucer, A. et al. Inhibition of cathepsin L lowers the apoptotic threshold of glioblastoma cells by up-regulating p53 and transcription of caspases 3 and 7. Apoptosis 16, 671–682 (2011). https://doi.org/10.1007/s10495-011-0600-6

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  • DOI: https://doi.org/10.1007/s10495-011-0600-6

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