Skip to main content

Advertisement

SpringerLink
Log in

Exogenous wt-p53 enhances the antitumor effect of HSV-TK/GCV on C6 glioma cells

  • Lab Investigation – Human/Animal Tissue
  • Published:
Journal of Neuro-Oncology Aims and scope Submit manuscript

Abstract

Objective

To study on the antitumor effect of combining wt-p53 gene with suicide gene therapy (HSV-tk+GCV) for malignant gliomas.

Methods

AdCMV-p53 was transfected into C6 glioma cells at MOI of (Multiplicity of infection) 0(G100), 10(TPG1), 100(TPG2), then AdCMV-tk was transducted to C6 glioma cells of G100, TPG1 and TPG2, respectively, at MOI of 100. The C6 glioma cells tranfected with both AdCMV-p53 and AdCMV-tk were exposed to various concentration of GCV. The cell survival rate was measured by MTT assay in vitro. Rat glioma model was established by injecting 5 × 105 C6 glioma cells into right caudate nucleus of SD rats. AdCMV-p53 and AdCMV-tk were injected into glioma on day 5 and 6, respectively. On day 7, ganciclovir (GCV) was administrated intraperitoneally at 15 mg/kg/day for 14 days. The survival time of all rats was observed. The growth of intracerebral tumors was monitored dynamically by enhanced MRI. Cell apoptosis was evaluated by TUNEL method. Expression of HSV-tk gene was identified by in situ hybridization and expression of exogenous p53 gene was detected with Western blotting.

Results

In vitro, wt-p53 significantly enhanced antitumor effect of HSV-tk/GCV. The concentration of GCV for ID50 of TPG2 cells (0.001 μg/ml GCV) was 10 times lower than that for the cells of tk-GCV group (MOI = 100), while the concentration of GCV for ID100 of TPG2 (0.01 μg/ml GCV) and TPG1(0.1 μg/ml GCV) was 100 and 10 times lower than that for the cells of tk-GCV group (MOI = 100), respectively. Apoptosis of C6 glioma cells also could be induced by transfection with wt-p53 gene slightly. For in vivo study, the survival time of tumor-bearing rats treated with HSV-TK/GCV or wt-p53 combined with HSV-TK/GCV was significantly prolonged and the intracerebral tumors were regressed and disappeared earlier in the combined gene therapy group than those in the HSV-TK/GCV therapy group as shown in enhanced MRI. However, only half dose of GCV for the rats treated with both wt-p53 and HSV-TK/GCV was needed to obtain the same efficacy as those rats treated with HSV-TK/GCV alone. These results indicate that the transfection of wt-p53 potentiates the effect of HSV-TK/GCV therapy.

Conclusions

The combination of HSV-tk/GCV system with wt-p53 gene transduction is optimal for clinical therapeutic trials of suicide gene therapy for malignant gliomas.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Moolten FL (1994) Drug sensitivity (bsuicideQ) genes for selective cancer chemotherapy. Cancer Gene Ther 1:279–287

    PubMed  CAS  Google Scholar 

  2. Hakumaki JM, Poptani H, Sandmair AM, Yla-Herttuala S, Kauppinen RA (1999) 1H MRS detects polyunsaturated fatty acid accumulation during gene therapy of glioma: implications for the in vivo detection of apoptosis. Nat Med 5:1323–1327

    Article  PubMed  CAS  Google Scholar 

  3. Rainov NGA (2000) Phase III clinical evaluation of herpes simplex virus type 1 thymidine kinase and ganciclovir gene therapy as an adjuvant to surgical resection and radiation in adults with previously untreated glioblastoma multiforme. Hum Gene Ther 11:2389–2401

    Article  PubMed  CAS  Google Scholar 

  4. Trask TW, Trask RP, Aguilar-Cordova E, Shine HD, Wyde PR, Goodman JC, Hamilton WJ, Rojas-Martinez A, Chen SH, Woo SL, Grossman RG (2000) Phase I study of adenoviral delivery of the HSV-tk gene and ganciclovir administration in patients with current malignant brain tumors. Mol Ther 1:195–203

    Article  PubMed  CAS  Google Scholar 

  5. Sandmair AM, Loimas S, Puranen P, Immonen A, Kossila M, Puranen M, Hurskainen H, Tyynela K, Turunen M, Vanninen R, Lehtolainen P, Paljarvi L, Johansson R, Vapalahti M, Yla-Herttuala S (2000) Thymidine kinase gene therapy for human malignant glioma, using replication-deficient retroviruses or adenoviruses. Hum Gene Ther 11:2197–2205

    Article  PubMed  CAS  Google Scholar 

  6. Immonen A, Vapalahti M, Tyynel K, Hurskainen H, Sandmair A, Vanninen R, Langford G, Murray N, Yla-Herttuala S (2004) AdvHSV-tk gene therapy with intravenous ganciclovir improves survival in human malignant glioma: a randomised, controlled study. Mol Ther 10:967–972

    Article  PubMed  CAS  Google Scholar 

  7. Namba H, Iwadate Y, Tagawa M, Kimura M, Shimizu H, Sato Y, Sueyoshi K, Sakiyama S (1996) Evaluation of the bystander effect in experiment brain tumors bearing HSV-TK gene by serial MRI. Hum Gene Ther 7:1847–1852

    PubMed  CAS  Google Scholar 

  8. Cool V, Pirotte B, Gerard C, Dargent JL, Baudson N, Levivier M, Goldman S, Hildebrand J, Brotchi J, Velu T (1996) Curative potenial of HSVtk gene transfer in rats with 9L gliosarcoma. Hum Gene Ther 7:627–635

    PubMed  CAS  Google Scholar 

  9. Di Meco F, Benedetti S, Colombo MP, Finocchiaro G (1997) Perspectives for the gene therapy of malignant gliomas by suicide gene transfer. J Neurosurgical Sci 41:227–234

    CAS  Google Scholar 

  10. Rosolen A, Frascella E, di Francesco C, Todesco A, Petrone M, Mehtali M, Zacchello F, Zanesco L, Scarpa M (1998) In vitro and in vivo antitumor effect of retrovirus-mediated herpes simplex thymidine kinase gene-transfer in human medulloblastoma. Gene Ther 5:113–120

    Article  PubMed  CAS  Google Scholar 

  11. Vincent AJ, Vogels R, Someren GV, Esandi MC, Noteboom JL, Avezaat CJ, Vecht C, Bekkum DW, Valerio D, Bout A, Hoogerbrugge PM (1996) Herpes simplex virus thymidine kinase gene therapy for rat malignant brain tumors. Hum Gene Ther 7:197–205

    PubMed  CAS  Google Scholar 

  12. Maron A, Gustin T, Le Roux A, Mottet I, Dedieu JF, Brion JP, Demeure R, Perricaudet M, Octave JN (1996) Gene therapy of rat C6 glioma using adenovirus-mediated transfer of the HSV-tk gene: long-term follow-up by magnetic resonance imaging. Gene Ther 3:315–322

    PubMed  CAS  Google Scholar 

  13. Perez-Cruet MJ, Trask TW, Chen SH, Goodman JC, Woo SL, Grossman RG, Shine HD (1994) Adenovirus-mediated gene therapy of experimental gliomas. J Neurosci Res 39:506–511

    Article  PubMed  CAS  Google Scholar 

  14. Smitt PS, Driesse M, Wolbers J, Kros M, Avezaat C (2003) Treatment of relapsed malignant glioma with an adenoviral vector containing the herpes simplex thymidine kinase gene followed by ganciclovir. Mol Ther 7:851–858

    Article  PubMed  CAS  Google Scholar 

  15. Symonds H, Krall L, Remington L, Saenz-Robles M, Lowe S, Jacks T, Dyke TV (1994) p53-dependent apoptosis suppresses tumor growth and progression in vivo. Cell 78:703–711

    Article  PubMed  CAS  Google Scholar 

  16. Niranjan A, Moriuchi S, Lunsford LD, Kondziolka D, Flickinger JC, Fellows W, Rajendiran S, Tamura M, Cohen JB, Glorioso JC (2000) Effective treatment of experimental glioblastoma by HSV verctor-mediated TNFα and HSV-tk gene transfer in combination with radiosurgery and ganciclovir administration. Mol Ther 2:114–120

    Article  PubMed  CAS  Google Scholar 

  17. Uckert W, Kammertons T, Haack K, Qin Z, Gebert J, Schendel DJ, Blankenstein T (1998) Double suicide gene (cytosine deaminase and herpes simplex virus thymidine kinase) but not single gene transfer allows reliable elimination of tumor cells in vivo. Hum Gene Ther 9:855–865

    Article  PubMed  CAS  Google Scholar 

  18. Terao S, Shirakawa T, Goda K, Kamidono S, Fujisawa M, Gotoh A (2005) Recombinant interleukin-2 enhanced the antitumor effect of ADV/RSV-HSV-tk/ACV therapy in a murine bladder cancer model. Anticancer Res 25:2757–2760

    PubMed  CAS  Google Scholar 

  19. Colombo F, Barzon L, Franchin E, Pacenti M, Pinna V, Danieli D, Zanusso M, Palu G (2005) Combined HSV-TK/IL-2 gene therapy in patients with recurrent glioblastoma multiforme: biological and clinical results. Cancer Gene Ther 12:835–848

    Article  PubMed  CAS  Google Scholar 

  20. Benedetti S, Dimeco F, Pollo B, Cirenei N, Colombo BM, Bruzzone MG, Cattaneo E, Vescovi A, Didonato S, Colombo MP, Finocchiaro G (1997) Limited efficacy of the HSVtk/GCV system for gene therapy of malignant gliomas and perspectives for the combined transduction of the IL-4 gene. Hum Gene Ther 8:1345–1353

    PubMed  CAS  Google Scholar 

  21. Beltinger C, Fulda S, Kammertoens T, Meyer E, Uckert W, Debatin KM (1999) Herpes simplex virus thymidine kinase/ganciclovir-induced apoptosis involves ligand-independent death receptor aggregation and activation of caspases. Proc Natl Acad Sci USA 96:8699–8704

    Article  PubMed  CAS  Google Scholar 

  22. Xie YJ, Gilbert JD, Kim JH, Freytag SO (1999) Efficacy of adenovirus-mediated CD/5-FC and HSV-1 thymidine kinase/ganciclovir suicide gene therapies concomitant with p53 gene therapy. Clinical Cancer Res 5:4224–4232

    CAS  Google Scholar 

  23. Fischer U, Steffens S, Frank S, Rainov NG, Schulze-Osthoff K, Kramm CM (2005) Mechanisms of thymidine kinase/ganciclovir and cytosine deaminase/5-fluorocytosine suicide gene therapy-induced cell death in glioma cells. Oncogene 24:1231–1243

    Article  PubMed  CAS  Google Scholar 

  24. Kock H, Harris MP, Anderson SC, Machemer T, Hancock W, Sutjipto S, Wills KN, Gregory RJ, Shepard HM, Westphal M, Maneval DC (1996) Adenovirus-mediated p53 gene transfer suppress growth of human glioblastoma cells in vitro and in vivo. Int J Cancer 67:808–815

    Article  PubMed  CAS  Google Scholar 

  25. Gomez-Manzano C, Fueyo J, Kyritsis AP, Steck PA, Roth JA, McDonnell TJ, Steck KD, Levin VA, Yung WK (1996) Adenovirus-mediated transfer of the p53 gene produces rapid and generalized death of human glioma cells via apoptosis. Cancer Res 56:694–699

    PubMed  CAS  Google Scholar 

  26. Harris MP, Sutjipto S, Wills KN, Hancock W, Cornell D, Johnson DE, Gregory RJ, Shepard HM, Maneval DC (1996) Adenovirus-mediated p53 gene transfer inhibits growth of human tumor cells expressing mutant p53 protein. Cancer Gene Ther 3:121–130

    PubMed  CAS  Google Scholar 

  27. Li H, Lochmuller H, Yong VW, Karpati G, Nalbantoglu J (1997) Adenovirus-mediated wild-type p53 gene transfer and overexpression induces apoptosis of human glioma cells independent of endogenous p53 status. J Neuropathol Exp Neurol 56:872–878

    PubMed  CAS  Google Scholar 

  28. Bi WL, Parysek LM, Warnick R, Stambrook PJ (1993) In vitro evidence that metabolic cooperation is responsible for the bystander effect observed with HSV-TK retroviral gene therapy. Hum Gene Ther 4:725–731

    PubMed  CAS  Google Scholar 

  29. Touraine RL, Ishii-Morita H, Ramsey WJ, Blaese RM (1998) The bystander effect in the HSVtk/ganciclovir system and its relation to gap junctional communication. Gene Ther 5:1705–1711

    Article  PubMed  CAS  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  31. Freeman SM, Abboud CN, Whartenby KA, Packman CH, Koeplin DS, Moolten FL, Abraham GN (1993) The bystander effect: tumor regression when a fraction of tumor mass is genetically modified. Cancer Res 53:5274–5283

    PubMed  CAS  Google Scholar 

  32. van Dillen IJ, Mulder NH, Sluiter WJ, Meijer C, de Jong S, Loncarek J, Mesnil M, de Vries EF, Vaalburg W, Hospers GA (2005) Influence of p53 status on the HSV-Tk/GCV-induced bystander effect in a panel of human ovarian carcinoma cell lines. Oncol Res 15:151–159

    PubMed  Google Scholar 

  33. Mishima K, Asai A, Sugiyama A, Miyagi Y, Kitanaka C, Kagaya S, Kirino T, Kuchino Y (1996) Increased expression of schwannoma-derived growth factor (SDGF) mRNA in rat tumor cells: involvement of SDGF in the growth promotion of rat gliomas. Int J Cancer 66:352–357

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Neurosurgery, Tianjin Medical University General Hospital, 154 Anshan Road, Tianjin, 300052, People’s Republic of China

    Qiang Huang, Peiyu Pu, Zhibo Xia & Yongping You

  2. Laboratory of Neuro-Oncology, Tianjin Neurological Institute, Tianjin, 300052, People’s Republic of China

    Qiang Huang, Peiyu Pu, Zhibo Xia & Yongping You

Authors
  1. Qiang Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peiyu Pu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhibo Xia
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yongping You
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Qiang Huang.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Huang, Q., Pu, P., Xia, Z. et al. Exogenous wt-p53 enhances the antitumor effect of HSV-TK/GCV on C6 glioma cells. J Neurooncol 82, 239–248 (2007). https://doi.org/10.1007/s11060-006-9279-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11060-006-9279-x

Keywords

Search

Navigation