Apoptosis induction with 5-fluorocytosine/cytosine deaminase gene therapy for human malignant glioma cells mediated by adenovirus

Kazuhiko Kurozumi, Takashi Tamiya, Yasuhiro Ono, Shinji Otsuka, Hirokazu Kambara, Yoshiaki Adachi, Tomotsugu Ichikawa, Hirofumi Hamada, Takashi Ohmoto

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Previously, we evaluated the therapeutic efficacy of the adenovirus-mediated transduction of the cytosine deaminase (CD) gene and 5-fluorocytosine (5-FC) for malignant gliomas. However, the molecular pathways that mediate the 5-FC/CD gene therapy-induced cell death remains to be elucidated. In this study, we examined the induction of apoptosis and the role of caspases in 5-FC/CD gene therapy using human malignant glioma cells [Gli36Δ5 (mutated p53) and U87MG (wild p53)]. The treatment with 5-FC/CD gene-therapy-induced apoptosis both in Gli36Δ5 cells and in U87MG cells according to flow cytometric analysis. Immunoblot analysis revealed that caspases 3 and 9 were processed in response to 5-FC/CD in a concentration- and time-dependent manner, but caspase 8 was not. Each caspase 3 and 9 inhibitor significantly reduced apoptosis triggered by 5-FC/CD, but the caspase 8 inhibitor did not affect apoptosis induction. 5-FC/CD significantly promoted the release of cytochorme c from mitochondria in a concentration-dependent manner. These results indicate that 5-FC/CD gene therapy induces apoptosis in human malignant glioma cells and that the apoptotic cell death is mediated by the activation of mitochondrial caspase cascades involving caspases 3 and 9. This is the first report concerning the apoptotic mechanism of 5-FC/CD gene therapy, and these findings could be used to increase the efficacy of suicide gene therapy systems for the treatment of malignant glioma.

Original languageEnglish
Pages (from-to)117-127
Number of pages11
JournalJournal of Neuro-Oncology
Volume66
Issue number1-2
DOIs
Publication statusPublished - Jan 2004

Fingerprint

Cytosine Deaminase
Flucytosine
Adenoviridae
Glioma
Genetic Therapy
Apoptosis
Caspase 9
Caspase 3
Caspase 8
Caspases
Cell Death
Caspase Inhibitors
Suicide
Mitochondria
Therapeutics

Keywords

  • 5-fluorocytosine
  • Adenovirus vector
  • Apoptosis
  • Caspase
  • Cytosine deaminase
  • Gene Therapy
  • Malignant gliomas
  • Mitochondria

ASJC Scopus subject areas

  • Clinical Neurology
  • Cancer Research
  • Oncology
  • Neuroscience(all)

Cite this

Apoptosis induction with 5-fluorocytosine/cytosine deaminase gene therapy for human malignant glioma cells mediated by adenovirus. / Kurozumi, Kazuhiko; Tamiya, Takashi; Ono, Yasuhiro; Otsuka, Shinji; Kambara, Hirokazu; Adachi, Yoshiaki; Ichikawa, Tomotsugu; Hamada, Hirofumi; Ohmoto, Takashi.

In: Journal of Neuro-Oncology, Vol. 66, No. 1-2, 01.2004, p. 117-127.

Research output: Contribution to journalArticle

Kurozumi, Kazuhiko ; Tamiya, Takashi ; Ono, Yasuhiro ; Otsuka, Shinji ; Kambara, Hirokazu ; Adachi, Yoshiaki ; Ichikawa, Tomotsugu ; Hamada, Hirofumi ; Ohmoto, Takashi. / Apoptosis induction with 5-fluorocytosine/cytosine deaminase gene therapy for human malignant glioma cells mediated by adenovirus. In: Journal of Neuro-Oncology. 2004 ; Vol. 66, No. 1-2. pp. 117-127.
@article{37349648eac749e584764b3ebb15b455,
title = "Apoptosis induction with 5-fluorocytosine/cytosine deaminase gene therapy for human malignant glioma cells mediated by adenovirus",
abstract = "Previously, we evaluated the therapeutic efficacy of the adenovirus-mediated transduction of the cytosine deaminase (CD) gene and 5-fluorocytosine (5-FC) for malignant gliomas. However, the molecular pathways that mediate the 5-FC/CD gene therapy-induced cell death remains to be elucidated. In this study, we examined the induction of apoptosis and the role of caspases in 5-FC/CD gene therapy using human malignant glioma cells [Gli36Δ5 (mutated p53) and U87MG (wild p53)]. The treatment with 5-FC/CD gene-therapy-induced apoptosis both in Gli36Δ5 cells and in U87MG cells according to flow cytometric analysis. Immunoblot analysis revealed that caspases 3 and 9 were processed in response to 5-FC/CD in a concentration- and time-dependent manner, but caspase 8 was not. Each caspase 3 and 9 inhibitor significantly reduced apoptosis triggered by 5-FC/CD, but the caspase 8 inhibitor did not affect apoptosis induction. 5-FC/CD significantly promoted the release of cytochorme c from mitochondria in a concentration-dependent manner. These results indicate that 5-FC/CD gene therapy induces apoptosis in human malignant glioma cells and that the apoptotic cell death is mediated by the activation of mitochondrial caspase cascades involving caspases 3 and 9. This is the first report concerning the apoptotic mechanism of 5-FC/CD gene therapy, and these findings could be used to increase the efficacy of suicide gene therapy systems for the treatment of malignant glioma.",
keywords = "5-fluorocytosine, Adenovirus vector, Apoptosis, Caspase, Cytosine deaminase, Gene Therapy, Malignant gliomas, Mitochondria",
author = "Kazuhiko Kurozumi and Takashi Tamiya and Yasuhiro Ono and Shinji Otsuka and Hirokazu Kambara and Yoshiaki Adachi and Tomotsugu Ichikawa and Hirofumi Hamada and Takashi Ohmoto",
year = "2004",
month = "1",
doi = "10.1023/B:NEON.0000013494.98345.80",
language = "English",
volume = "66",
pages = "117--127",
journal = "Journal of Neuro-Oncology",
issn = "0167-594X",
publisher = "Kluwer Academic Publishers",
number = "1-2",

}

TY - JOUR

T1 - Apoptosis induction with 5-fluorocytosine/cytosine deaminase gene therapy for human malignant glioma cells mediated by adenovirus

AU - Kurozumi, Kazuhiko

AU - Tamiya, Takashi

AU - Ono, Yasuhiro

AU - Otsuka, Shinji

AU - Kambara, Hirokazu

AU - Adachi, Yoshiaki

AU - Ichikawa, Tomotsugu

AU - Hamada, Hirofumi

AU - Ohmoto, Takashi

PY - 2004/1

Y1 - 2004/1

N2 - Previously, we evaluated the therapeutic efficacy of the adenovirus-mediated transduction of the cytosine deaminase (CD) gene and 5-fluorocytosine (5-FC) for malignant gliomas. However, the molecular pathways that mediate the 5-FC/CD gene therapy-induced cell death remains to be elucidated. In this study, we examined the induction of apoptosis and the role of caspases in 5-FC/CD gene therapy using human malignant glioma cells [Gli36Δ5 (mutated p53) and U87MG (wild p53)]. The treatment with 5-FC/CD gene-therapy-induced apoptosis both in Gli36Δ5 cells and in U87MG cells according to flow cytometric analysis. Immunoblot analysis revealed that caspases 3 and 9 were processed in response to 5-FC/CD in a concentration- and time-dependent manner, but caspase 8 was not. Each caspase 3 and 9 inhibitor significantly reduced apoptosis triggered by 5-FC/CD, but the caspase 8 inhibitor did not affect apoptosis induction. 5-FC/CD significantly promoted the release of cytochorme c from mitochondria in a concentration-dependent manner. These results indicate that 5-FC/CD gene therapy induces apoptosis in human malignant glioma cells and that the apoptotic cell death is mediated by the activation of mitochondrial caspase cascades involving caspases 3 and 9. This is the first report concerning the apoptotic mechanism of 5-FC/CD gene therapy, and these findings could be used to increase the efficacy of suicide gene therapy systems for the treatment of malignant glioma.

AB - Previously, we evaluated the therapeutic efficacy of the adenovirus-mediated transduction of the cytosine deaminase (CD) gene and 5-fluorocytosine (5-FC) for malignant gliomas. However, the molecular pathways that mediate the 5-FC/CD gene therapy-induced cell death remains to be elucidated. In this study, we examined the induction of apoptosis and the role of caspases in 5-FC/CD gene therapy using human malignant glioma cells [Gli36Δ5 (mutated p53) and U87MG (wild p53)]. The treatment with 5-FC/CD gene-therapy-induced apoptosis both in Gli36Δ5 cells and in U87MG cells according to flow cytometric analysis. Immunoblot analysis revealed that caspases 3 and 9 were processed in response to 5-FC/CD in a concentration- and time-dependent manner, but caspase 8 was not. Each caspase 3 and 9 inhibitor significantly reduced apoptosis triggered by 5-FC/CD, but the caspase 8 inhibitor did not affect apoptosis induction. 5-FC/CD significantly promoted the release of cytochorme c from mitochondria in a concentration-dependent manner. These results indicate that 5-FC/CD gene therapy induces apoptosis in human malignant glioma cells and that the apoptotic cell death is mediated by the activation of mitochondrial caspase cascades involving caspases 3 and 9. This is the first report concerning the apoptotic mechanism of 5-FC/CD gene therapy, and these findings could be used to increase the efficacy of suicide gene therapy systems for the treatment of malignant glioma.

KW - 5-fluorocytosine

KW - Adenovirus vector

KW - Apoptosis

KW - Caspase

KW - Cytosine deaminase

KW - Gene Therapy

KW - Malignant gliomas

KW - Mitochondria

UR - http://www.scopus.com/inward/record.url?scp=1342308405&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=1342308405&partnerID=8YFLogxK

U2 - 10.1023/B:NEON.0000013494.98345.80

DO - 10.1023/B:NEON.0000013494.98345.80

M3 - Article

C2 - 15015777

AN - SCOPUS:1342308405

VL - 66

SP - 117

EP - 127

JO - Journal of Neuro-Oncology

JF - Journal of Neuro-Oncology

SN - 0167-594X

IS - 1-2

ER -