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Connexin 43–mediated bystander effect in two rat glioma cell models

Cancer Gene Therapy volume 9, pages 149–155 (2002)Cite this article

Abstract

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-α-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.

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References

  1. Nagy JI, Rash JE . Connexins and gap junctions of astrocytes and oligodendrocytes in the CNS Brain Res Brain Res Rev 2000 32: 29–44

    Article  CAS  Google Scholar 

  2. Kumar NM, Gilula NB . The gap junction communication channel Cell 1996 84: 381–388

    Article  CAS  Google Scholar 

  3. Culver KW, Ram Z, Wallbridge S, Ishii H, Oldfield EH, Blaese RM . In vivo gene transfer with retroviral vector-producer cells for treatment of experimental brain tumors Science 1992 256: 1550–1552

    Article  CAS  Google Scholar 

  4. Fick J, Barker FG, Dazin P, Westphale EM, Beyer EC, Israel MA . The extent of heterocellular communication mediated by gap junctions is predictive of bystander tumor cytotoxicity in vitro Proc Natl Acad Sci USA 1995 92: 11071–11075

    Article  CAS  Google Scholar 

  5. Elshami AA, Saavedra A, Zhang H et al. Gap junctions play a role in the “bystander effect” of the herpes simplex virus thymidine kinase/ganciclovir system in vitro Gene Ther 1996 3: 85–92

    CAS  PubMed  Google Scholar 

  6. Mesnil M, Piccoli C, Tiraby G, Willecke K, Yamasaki H . Bystander killing of cancer cells by herpes simplex virus thymidine kinase gene is mediated by connexins Proc Natl Acad Sci USA 1996 93: 1831–1835

    Article  CAS  Google Scholar 

  7. Vrionis FD, Wu JK, Qi P, Waltzman M, Cherington V, Spray DC . The bystander effect exerted by tumor cells expressing the herpes simplex virus thymidine kinase (HSVTK) gene is dependent on connexin expression and cell communication via gap junctions Gene Ther 1997 4: 577–585

    Article  CAS  Google Scholar 

  8. Kruse CA, Molleston MC, Parks EP, Schiltz PM, Kleinschmidt-De Masters BK, Hickey WF . A rat glioma model, CNS-1, with invasive characteristics similar to those of human gliomas: a comparison to 9L gliosarcoma J Neuro-Oncol 1994 22: 191–200

    Article  CAS  Google Scholar 

  9. Boussif O, Lezoualc'h F, Zanta MA et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine Proc Natl Acad Sci USA 1995 92: 7297–7301

    Article  CAS  Google Scholar 

  10. Mosmann T . Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays J Immunol Methods 1983 65: 55–63

    Article  CAS  Google Scholar 

  11. Davidson JS, Baumgarten IM, Harley EH . Reversible inhibition of intercellular junctional communication by glycyrrhetinic acid Biochem Biophys Res Commun 1986 134: 29–36

    Article  CAS  Google Scholar 

  12. Chen CY, Chang YN, Ryan P, Linscott M, McGarrity GJ, Chiang YL . Effect of herpes simplex virus thymidine kinase expression levels on ganciclovir-mediated cytotoxicity and the “bystander effect.” Hum Gene Ther 1995 6: 1467–1476

    Article  CAS  Google Scholar 

  13. Goldberg GS, Bechberger JF, Naus CCG . A pre-loading method of evaluating gap junctional communication by fluorescent dye transfer Biotechniques 1995 18: 210–217

    Google Scholar 

  14. Ram Z, Culver KW, Walbridge S, Blaese RM, Oldsfield EH . In situ retroviral-mediated gene transfer for the treatment of brain tumors in rats Cancer Res 1993 53: 83–88

    CAS  PubMed  Google Scholar 

  15. Freeman SM, Abboud CN, Whartenby KA et al. The “bystander effect”: tumor regression when a fraction of the tumor mass is genetically modified Cancer Res 1993 53: 5274–5283

    CAS  Google Scholar 

  16. Sturtz FG, Wadell K, Shulok J et al. Gene therapy for glioblastoma: parameters involved in the {NdT: à vérifier. Strictement ne doit pas avoit le “the”} thymidine kinase/ganciclovir-mediated killing Hum Gene Ther 1997 8: 1945–1953

    Article  CAS  Google Scholar 

  17. Bi W, Parysek LM, Warmick R, Stambrook PJ . In vitro evidence that metabolic cooperation is responsible for the bystander effect observed with HSVtk retroviral gene therapy Hum Gene Ther 1993 4: 725–731

    Article  CAS  Google Scholar 

  18. Samejima Y, Meruelo D . Bystander killing induces apoptosis and is inhibited by forskolin Gene ther 1995 2: 50–58

    CAS  PubMed  Google Scholar 

  19. Rubsam LZ, Boucher PD, Murphy PJ, Kukuruga M, Shewach DS . Cytotoxicity and accumulation of ganciclovir triphosphate in bystander cells cocultured with herpes simplex virus type 1 thymidine kinase–expressing human glioblastoma cells Cancer Res 1999 59: 669–675

    CAS  PubMed  Google Scholar 

  20. Dilber MS, Abedi MR, Christensson B et al. Gap junctions promote the bystander effect of herpes simplex virus thymidine kinase in vivo Cancer Res 1997 57: 1523–1528

    CAS  PubMed  Google Scholar 

  21. Li H, Liu TF, Lazrak A et al. Properties and regulation of gap junctional hemichannels in the plasma membranes of cultured cells J Cell Biol 1996 134: 1019–1030

    Article  CAS  Google Scholar 

  22. Robinson SR, Hampson ECGM, Munro MN, Vaney DI . Unidirectional coupling of gap junctions between neuroglia Science 1993 262: 1072–1074

    Article  CAS  Google Scholar 

  23. Lin JHC, Weigel H, Cotrina ML et al. Gap-junction–mediated propagation and amplification of cell injury Nat Neurosci 1998 1: 494–500

    Article  CAS  Google Scholar 

  24. Cinerei N, Colombo BM, Mesnil M, Benedetti S, Yamasaki H, Finocchiaro G . In vitro and in vivo effects of retrovirus-mediated transfer of the connexin 43 gene in malignant gliomas: consequences for HSVtk/GCV anticancer gene therapy Gene Ther 1998 5: 1221–1226

    Article  Google Scholar 

  25. Touraine RL, Vahanian N, Ramsey WJ, Blaese RM . Enhancement of the herpes simplex virus thymidine kinase/ganciclovir bystander effect and its antitumor efficacy in vivo by pharmacological manipulation of gap junctions Hum Gen Ther 1998 9: 2385–2391

    Article  CAS  Google Scholar 

  26. Zhu W, Mironov N, Yamasaki H . Increased genetic stability of HeLa cells after connexin 43 gene transfection Cancer 1997 57: 2148–2150

    CAS  Google Scholar 

  27. Zhu D, Caveney S, Kidder GM, Naus CCG . Transfection of C6 glioma cells with connexin 43 cDNA: analysis of expression, intercellular coupling, and cell proliferation Proc Natl Acad Sci USA 1991 88: 1883–1887

    Article  CAS  Google Scholar 

  28. Naus CCG, Elisevich K, Zhu D, Belliveau DJ, Del Maestro RF . In vivo growth of C6 glioma cells transfected with connexin 43 cDNA Cancer Res 1992 52: 4208–4213

    CAS  PubMed  Google Scholar 

  29. Lau AF, Kurata WE, Kanemitsu MY et al. Regulation of connexin 43 function by activated tyrosine protein kinase J Bioenerg Biomembr 1996 28: 359–368

    Article  CAS  Google Scholar 

  30. Estin D, Li M, Spray D, Wu JK . Connexins are expressed in primary brain tumors and enhance the bystander effect in gene therapy Neurosurgery 1999 44: 361–368

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the Ligue Nationale contre le Cancer, the Association pour la Recherche sur les tumeurs cérébrales, and the AP-HP/CANAM.

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Authors and Affiliations

  1. Biologie des interactions neurones-glie, Unité INSERM U495, Université P et M Curie Hôpital de la Salpêtrière, Paris, France

    Marc Sanson, Véronique Marcaud, Eric Robin & Bernard Zalc

  2. Department of Neurosurgery, Hôpital de la Salpêtrière, Paris, France

    Charles Valéry

  3. Department of Oncology, Hôpital Boucicaut, Paris, France

    Franck Sturtz

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  1. Marc Sanson
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Correspondence to Marc Sanson.

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Sanson, M., Marcaud, V., Robin, E. et al. Connexin 43–mediated bystander effect in two rat glioma cell models. Cancer Gene Ther 9, 149–155 (2002). https://doi.org/10.1038/sj.cgt.7700411

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