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

Armen Azatian, Hong Yu, Wande Dai, Fiona Schneiders, Natalia K. Botelho, Reginald V. N. Lord

Research output: Contribution to journalArticle

17 Citations (Scopus)

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 microg/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 junand 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.
Original languageEnglish
Pages (from-to)1044-1051
Number of pages8
JournalJournal of Gastrointestinal Surgery
Volume13
Issue number6
DOIs
Publication statusPublished - 2009

Fingerprint

Esophagogastric Junction
Genetic Therapy
Ganciclovir
Suicide
Stomach
Adenocarcinoma
Terminal Repeat Sequences
Neoplasms
Cell Survival
Starvation
Nude Mice
Cell Line
Genes
Glucose
Murine Leukemia Viruses
Thymidine Kinase
Prodrugs
Simplexvirus
Intraperitoneal Injections
Reverse Transcription

Cite this

Azatian, Armen ; Yu, Hong ; Dai, Wande ; Schneiders, Fiona ; Botelho, Natalia K. ; Lord, Reginald V. N. / Effectiveness of HSV-tk Suicide Gene Therapy Driven by the Grp78 Stress-inducible Promoter in Esophagogastric Junction and Gastric Adenocarcinomas. In: Journal of Gastrointestinal Surgery. 2009 ; Vol. 13, No. 6. pp. 1044-1051.
@article{bbb9fa44fc614490b558c00fb96060db,
title = "Effectiveness of HSV-tk Suicide Gene Therapy Driven by the Grp78 Stress-inducible Promoter in Esophagogastric Junction and Gastric Adenocarcinomas.",
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 microg/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 junand 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.",
keywords = "Gastric adenocarcinoma, Gastric neoplasms, Gastroesophageal junction adenocarcinoma, Gene therapy, Grp78, HSV-tk",
author = "Armen Azatian and Hong Yu and Wande Dai and Fiona Schneiders and Botelho, {Natalia K.} and Lord, {Reginald V. N.}",
note = "Imported on 12 Apr 2017 - DigiTool details were: Journal title (773t) = Journal of Gastrointestinal Surgery. ISSNs: 1091-255X;",
year = "2009",
doi = "10.1007/s11605-009-0839-1",
language = "English",
volume = "13",
pages = "1044--1051",
journal = "Journal of Gastrointestinal Surgery",
issn = "1091-255X",
publisher = "Springer",
number = "6",

}

Effectiveness of HSV-tk Suicide Gene Therapy Driven by the Grp78 Stress-inducible Promoter in Esophagogastric Junction and Gastric Adenocarcinomas. / Azatian, Armen; Yu, Hong; Dai, Wande; Schneiders, Fiona; Botelho, Natalia K.; Lord, Reginald V. N.

In: Journal of Gastrointestinal Surgery, Vol. 13, No. 6, 2009, p. 1044-1051.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Effectiveness of HSV-tk Suicide Gene Therapy Driven by the Grp78 Stress-inducible Promoter in Esophagogastric Junction and Gastric Adenocarcinomas.

AU - Azatian, Armen

AU - Yu, Hong

AU - Dai, Wande

AU - Schneiders, Fiona

AU - Botelho, Natalia K.

AU - Lord, Reginald V. N.

N1 - Imported on 12 Apr 2017 - DigiTool details were: Journal title (773t) = Journal of Gastrointestinal Surgery. ISSNs: 1091-255X;

PY - 2009

Y1 - 2009

N2 - 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 microg/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 junand 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.

AB - 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 microg/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 junand 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.

KW - Gastric adenocarcinoma

KW - Gastric neoplasms

KW - Gastroesophageal junction adenocarcinoma

KW - Gene therapy

KW - Grp78

KW - HSV-tk

U2 - 10.1007/s11605-009-0839-1

DO - 10.1007/s11605-009-0839-1

M3 - Article

VL - 13

SP - 1044

EP - 1051

JO - Journal of Gastrointestinal Surgery

JF - Journal of Gastrointestinal Surgery

SN - 1091-255X

IS - 6

ER -