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Effectiveness of HSV-tk Suicide Gene Therapy Driven by the Grp78 Stress-Inducible Promoter in Esophagogastric Junction and Gastric Adenocarcinomas

  • Original Article
  • Published:
Journal of Gastrointestinal Surgery

Abstract

Background

The thymidine kinase gene of the herpes simplex virus (HSV-tk) is a suicide gene when administrated with the prodrug ganciclovir (GCV). This study investigated the effectiveness of HSV-tk activation as gene therapy for gastroesophageal junction and gastric adenocarcinomas using either the stress-inducible Grp78 promoter or the murine leukemia virus long-terminal repeat (LTR) promoter.

Methods

The HSV-tk gene, controlled by either the Grp78 promoter or the LTR promoter, was transduced into the gastroesophageal junction adenocarcinoma cell line SK-GT-5 and the gastric adenocarcinoma cell line MKN-74. Cell viability after exposure to varying concentrations of GCV was compared. The same cell lines were used to develop a nude mouse model for studies of the HSV-tk/GCV effect in vivo. The effect of intraperitoneal GCV injection on growth of the subcutaneous tumors was measured. HSV-TK expression was measured by Western blot and reverse transcription polymerase chain reaction.

Results

Cell viability in vitro was significantly lower in the HSV-tk expressing (HSV-tk+) cells compared to control (no HSV-tk) cells after exposure to GCV. MKN-74tk+ cells were more sensitive to GCV killing than SK-GT-5tk+ cells. After culture with 1 μg/ml GCV for 10 days, MKN-74/tk cells were totally killed, whereas most SK-GT-5/tk cells survived. Cell viability was significantly lower under glucose starvation conditions when HSV-tk expression was regulated by the Grp78 promoter compared with the LTR promoter. MKN-74 tumors formed with HSV-tk+ cells in nude mice were eliminated after administration of GCV for 3 weeks, but GCV had no effect on tumors formed from HSV-tk− cells. Eradication of tumor formed with Grp78-tk cells was faster than that with LTR-tk cells. HSV-TK protein and mRNA were expressed in the transduced, but not the non-transduced tumors.

Conclusion

HSV-tk xwith ganciclovir suicide gene therapy results in significant cell killing in gastroesophageal junction and gastric adenocarcinoma cells both in vitro and in vivo, but complete tumor elimination only occurred with the gastric adenocarcinoma cell tumors. The most effective approach in this study used the Grp78 promoter in glucose-starvation stress conditions.

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Acknowledgment

This study is supported by STOP Cancer Foundation, Los Angeles, Cancer Institute NSW, NSW Cancer Council, and the National Health and Medical Research Council.

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Author notes

  1. Hong Yu

    Present address: Vascular Biology Institute, University of Miami, and Bruce W. Carter Miami Veterances Affairs Medical Center, 1201 NW 16th St, Miami, FL, 33125, USA

Authors and Affiliations

  1. Department of Surgery, University of Southern California Keck School of Medicine, Los Angeles, CA, 90033, USA

    Armen Azatian, Hong Yu, Wande Dai & Reginald V. N. Lord

  2. St. Vincent’s Centre for Applied Medical Research and Department of Surgery, St. Vincent’s Hospital, University of New South Wales, Sydney, 2010, Australia

    Fiona I. Schneiders, Natalia K. Botelho & Reginald V. N. Lord

  3. Suite 606, St. Vincent’s Clinic, 438 Victoria Street, Darlinghurst, Sydney, NSW, 2010, Australia

    Reginald V. N. Lord

Authors
  1. Armen Azatian
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  2. Hong Yu
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  3. Wande Dai
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  4. Fiona I. Schneiders
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  6. Reginald V. N. Lord
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Corresponding author

Correspondence to Reginald V. N. Lord.

Additional information

Armen Azatian and Hong Yu are equal co-first authors.

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Azatian, A., Yu, H., Dai, W. et al. Effectiveness of HSV-tk Suicide Gene Therapy Driven by the Grp78 Stress-Inducible Promoter in Esophagogastric Junction and Gastric Adenocarcinomas. J Gastrointest Surg 13, 1044–1051 (2009). https://doi.org/10.1007/s11605-009-0839-1

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  • DOI: https://doi.org/10.1007/s11605-009-0839-1

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