Connexin 43-mediated bystander effect in two rat glioma cell models.
In tumor models, the killing by ganciclovir of a fraction of tumor cells transfected with the thymidine kinase (TK) gene has been shown to induce complete regression of the tumor. The mechanism responsible for this bystander effect is thought to be the diffusion of toxic metabolites or apoptotic signals across gap junctions. Connexin 43 (Cx43) is the major component of astrocyte gap junctions. We investigated the susceptibility of two rat glioma cell lines (CNS1 and C6) to thymidine kinase/ganciclovir, before and after transfection with the Cx43 gene. We report a close correlation between the level of Cx43 expression, the extent of gap junctional communication and the amplitude of the bystander effect. Transfection of C6 cells (which display a weak bystander effect and low levels of connexin) with a Cx43 construct induced a strong bystander effect. Inhibition of gap junction activity by 18-alpha-glycyrrhetinic acid abolished the metabolic interaction between TK(+) and TK(-) cells. This metabolic interaction was also abolished if TK(+) and TK(-) cells were separated by a semipermeable membrane. Surprisingly, the transfection of only one of these two interacting cell types with the Cx43 gene was sufficient to induce a bystander effect, although this effect was weaker than that observed if both TK(+) and TK(-) cells expressed Cx43. Finally, Cx43 expression increased sensitivity to contact inhibition. Overall, our data provide evidence that the restoration of gap junctional communication may potentiate HSV/tk-based cancer treatment and suggest that this strategy may have wider application in cancer therapy.
Sanson
Marc
M
Biologie des interactions neurones-glie, Unité INSERM U495, Université P et M Curie Hôpital de la Salpêtrière, Paris, France. sanson@ccr.jussieu.fr
Marcaud
Véronique
V
Robin
Eric
E
Valéry
Charles
C
Sturtz
Franck
F
Zalc
Bernard
B
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D001932:Brain Neoplasms / Q000378:metabolism / Q000628:therapy*; D024201:Bystander Effect; D016923:Cell Death / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D018031:Connexin 43 / Q000235:genetics / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D015774:Ganciclovir / Q000494:pharmacology; D017629:Gap Junctions / Q000378:metabolism; D015316:Genetic Therapy; D005910:Glioma / Q000378:metabolism / Q000628:therapy*; D051381:Rats; D018139:Simplexvirus / Q000201:enzymology; D013937:Thymidine Kinase / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism
D000998:Antiviral Agents; D018031:Connexin 43; D013937:Thymidine Kinase; D015774:Ganciclovir