Induction of apoptosis in human esophageal cancer cells by sequential transfer of the wild-type p53 and E2F-1 genes: Involvement of p53 accumulation via ARF-mediated MDM2 down-regulation

Takahiro Itoshima, Toshiyoshi Fujiwara, Toshihiko Waku, Jianghua Shao, Masafumi Kataoka, Wendell G. Yarbrough, Ta Jen Liu, Jack A. Roth, Noriaki Tanaka, Masashi Kodama

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Transcriptional factor E2F-1 as well as tumor suppressor p53 have been shown to cause apoptosis independently in some types of human cancer cells when overexpressed. Here we report that sequential transfer of the wild-type p53 and E2F-1 genes efficiently induces apoptosis in human esophageal cancer cells and that E2F-1 overexpression directly activates expression of p14 (ARF), which inhibits MDM2-mediated p53 degradation, resulting in the stabilization of p53. Infection of human esophageal cancer cell lines T.Tn and TE8 with adenovirus vector-expressing E2F-1 (ad-E2F-1) enhanced mRNA and protein expression of ARF and decreased MDM2 protein expression. Transfection of ARF plasmid decreased MDM2 protein expression, which in turn increased p53 protein expression. Infection of T.Tn and TE8 cells first with adenovirus- expressing wild-type p53 (Ad-p53) and then with Ad-E2F-1 resulted in rapid induction of apoptosis; in contrast, simultaneous infection with Ad-E2F-1 and Ad-p53 had no significant antitumor effect. As shown by Western blot analysis, infection with suboptimal concentrations of Ad-E2F-1 induced the accumulation of exogenous p53 transduced by suboptimal concentrations of Ad- p53. Moreover, Ad-E2F-1-mediated ARF expression inhibited the up-regulation of MDM2 by overexpressed p53 in TE8 cells. Thus, overexpression of ectopic E2F-1 protein may stabilize endogenous as well as ectopic p53 protein via the E2F-1/ARF/MDM2/p53 regulatory pathway and, in this way, render cells more sensitive to apoptosis, an outcome that has important implications for the treatment of human esophageal cancer.

Original languageEnglish
Pages (from-to)2851-2859
Number of pages9
JournalClinical Cancer Research
Volume6
Issue number7
Publication statusPublished - Jul 2000

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p53 Genes
Esophageal Neoplasms
Down-Regulation
Apoptosis
E2F Transcription Factors
Infection
Adenoviridae
Proteins
Tumor Suppressor Protein p14ARF
Transfection
Neoplasms
Plasmids
Up-Regulation
Western Blotting
Cell Line
Messenger RNA
Genes

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Induction of apoptosis in human esophageal cancer cells by sequential transfer of the wild-type p53 and E2F-1 genes : Involvement of p53 accumulation via ARF-mediated MDM2 down-regulation. / Itoshima, Takahiro; Fujiwara, Toshiyoshi; Waku, Toshihiko; Shao, Jianghua; Kataoka, Masafumi; Yarbrough, Wendell G.; Liu, Ta Jen; Roth, Jack A.; Tanaka, Noriaki; Kodama, Masashi.

In: Clinical Cancer Research, Vol. 6, No. 7, 07.2000, p. 2851-2859.

Research output: Contribution to journalArticle

Itoshima, Takahiro ; Fujiwara, Toshiyoshi ; Waku, Toshihiko ; Shao, Jianghua ; Kataoka, Masafumi ; Yarbrough, Wendell G. ; Liu, Ta Jen ; Roth, Jack A. ; Tanaka, Noriaki ; Kodama, Masashi. / Induction of apoptosis in human esophageal cancer cells by sequential transfer of the wild-type p53 and E2F-1 genes : Involvement of p53 accumulation via ARF-mediated MDM2 down-regulation. In: Clinical Cancer Research. 2000 ; Vol. 6, No. 7. pp. 2851-2859.
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abstract = "Transcriptional factor E2F-1 as well as tumor suppressor p53 have been shown to cause apoptosis independently in some types of human cancer cells when overexpressed. Here we report that sequential transfer of the wild-type p53 and E2F-1 genes efficiently induces apoptosis in human esophageal cancer cells and that E2F-1 overexpression directly activates expression of p14 (ARF), which inhibits MDM2-mediated p53 degradation, resulting in the stabilization of p53. Infection of human esophageal cancer cell lines T.Tn and TE8 with adenovirus vector-expressing E2F-1 (ad-E2F-1) enhanced mRNA and protein expression of ARF and decreased MDM2 protein expression. Transfection of ARF plasmid decreased MDM2 protein expression, which in turn increased p53 protein expression. Infection of T.Tn and TE8 cells first with adenovirus- expressing wild-type p53 (Ad-p53) and then with Ad-E2F-1 resulted in rapid induction of apoptosis; in contrast, simultaneous infection with Ad-E2F-1 and Ad-p53 had no significant antitumor effect. As shown by Western blot analysis, infection with suboptimal concentrations of Ad-E2F-1 induced the accumulation of exogenous p53 transduced by suboptimal concentrations of Ad- p53. Moreover, Ad-E2F-1-mediated ARF expression inhibited the up-regulation of MDM2 by overexpressed p53 in TE8 cells. Thus, overexpression of ectopic E2F-1 protein may stabilize endogenous as well as ectopic p53 protein via the E2F-1/ARF/MDM2/p53 regulatory pathway and, in this way, render cells more sensitive to apoptosis, an outcome that has important implications for the treatment of human esophageal cancer.",
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AU - Shao, Jianghua

AU - Kataoka, Masafumi

AU - Yarbrough, Wendell G.

AU - Liu, Ta Jen

AU - Roth, Jack A.

AU - Tanaka, Noriaki

AU - Kodama, Masashi

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