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Bystander effect in glioblastoma cells with a predominant cytoplasmic localization of connexin43

Cancer Gene Therapy volume 15, pages 823–831 (2008)Cite this article

Abstract

Herpes simplex virus thymidine kinase (TK) gene transfer followed by ganciclovir (GCV) administration is an approach investigated for glioblastoma treatment. The bystander effect (BE) enhances the cytotoxic effect of this strategy by allowing the diffusion of phosphorylated GCV from TK-expressing cells toward neighboring TK negative cells. This transfer of toxic metabolites is mainly mediated via gap junctions that are composed of connexins. Downregulation and/or cytoplasmic localization of connexins are common in tumors, and should be detrimental to the success of the TK/GCV strategy. In this study, we investigated the level of expression, the localization and the functionality of connexin43 (Cx43) in three glioblastoma cell lines. We showed that Cx43 was predominantly located in lysosomes and late endosomes, with only few gap junctions present at the cell surface. Surprisingly, the gap-junctional intercellular communication (GJIC) and the BE capacity were preserved, and in two of the cell lines analyzed, it was at least twice as high as compared to a control HeLa transfectant that expresses high levels of Cx43 at the cell membrane. Experiments performed in the presence of α-glycyrrhetinic acid or small interfering RNA confirmed that Cx43 was responsible for the GJIC and the BE. Our results indicate for the first time that the very limited numbers of gap junctions present in glioblastoma cells are highly functional. We thus conclude that the TK/GCV strategy is still a valuable therapeutic option to be developed for the treatment of glioblastoma patients.

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Acknowledgements

We are grateful to Pedro Otavio de Campos-Lima for critical reading of the manuscript and to Alan Lau for providing the rat Cx43 cDNA used to generate HeLa/Cx43 cells. This study was initiated with a grant from the Canadian Institute of Health Research (CIHR) (IC074582). MC is a Senior Research Scholar of the Fonds de la Recherche en Santé du Québec (FRSQ) and during the course of this study SC held a graduate student award from the MCETC/CIHR strategic training program.

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  1. Centre de Recherche en Cancérologie de l’Université Laval, L’Hôtel-Dieu de Québec, Centre Hospitalier Universitaire de Québec, Québec, Canada

    S Cottin, K Ghani & M Caruso

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  1. S Cottin
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  2. K Ghani
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  3. M Caruso
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Correspondence to M Caruso.

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Cottin, S., Ghani, K. & Caruso, M. Bystander effect in glioblastoma cells with a predominant cytoplasmic localization of connexin43. Cancer Gene Ther 15, 823–831 (2008). https://doi.org/10.1038/cgt.2008.49

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