Level of reactive oxygen species induced by p21WAF(1)/CIP(1) is critical for the determination of cell fate

Takafumi Inoue, Kiyoko Kato, Hidenori Kato, Kazuo Asanoma, Ayumi Kuboyama, Yousuke Ueoka, Shinichiro Yamaguchi, Tatsuhiro Ogami, Norio Wake

Research output: Contribution to journalArticle

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Abstract

p21WAF(1)/CIP(1) is a well-known cell cycle regulatory protein which is overexpressed in several cancer cell lines, and known to determine cell fate. We generated three recombinant adenovirus vectors that expressed either the full-length p21 (Ad-p21F), a p21 mutant with a deletion of the C-terminal proliferative cell nuclear antigen (PCNA) binding domain (Ad-p21N), or a p21 mutant with a deletion of the N-terminal cyclin-dependent kinase binding domain (Ad-p21C). We transfected these vectors into five cancer cell lines. Premature senescence was induced in all of the lines only following transfection with Ad-p21N and Ad-p21F. In addition, apoptosis was also induced in LoVo and HCT116 cells that harbored wild-type p53 and the reactive oxygen species (ROS) level was higher than in senescent cells. Finally, the induction of apoptosis was inhibited by using siRNA to downregulate p53. This observation implies that there is a feedback signaling loop involving p21/ROS/p53 in apoptotic responses. It appears to be, at least in part, driven by high levels of p21 protein. Next, we investigated the cell death effect of endogenous p21 protein on cell fate using sodium butyrate (NaB). Treatment with 1 mM NaB or 2 to 5 mM NaB induced senescence or apoptosis, respectively. The level of intracellular ROS in 5 mM NaB treated cells was 2-fold higher, compared with that in 1 mM NaB treated cells. We also demonstrated that DNA damage response signals including ataxia telangiectasia mutated, γH2AX, and p38.MAPK were involved in NaB-induced cell death. The magnitude of intracellular ROS levels in response to p21 elicited either senescence or apoptosis in the cancer cell lines.

Original languageEnglish
Pages (from-to)1275-1283
Number of pages9
JournalCancer Science
Volume100
Issue number7
DOIs
Publication statusPublished - Jun 23 2009

Fingerprint

Reactive Oxygen Species
Apoptosis
Cell Line
Cell Death
HCT116 Cells
Ataxia Telangiectasia
Neoplasms
Nuclear Antigens
Cell Cycle Proteins
Butyric Acid
Cyclin-Dependent Kinases
Adenoviridae
Small Interfering RNA
DNA Damage
Transfection
Proteins
Down-Regulation

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

Level of reactive oxygen species induced by p21WAF(1)/CIP(1) is critical for the determination of cell fate. / Inoue, Takafumi; Kato, Kiyoko; Kato, Hidenori; Asanoma, Kazuo; Kuboyama, Ayumi; Ueoka, Yousuke; Yamaguchi, Shinichiro; Ogami, Tatsuhiro; Wake, Norio.

In: Cancer Science, Vol. 100, No. 7, 23.06.2009, p. 1275-1283.

Research output: Contribution to journalArticle

Inoue, Takafumi ; Kato, Kiyoko ; Kato, Hidenori ; Asanoma, Kazuo ; Kuboyama, Ayumi ; Ueoka, Yousuke ; Yamaguchi, Shinichiro ; Ogami, Tatsuhiro ; Wake, Norio. / Level of reactive oxygen species induced by p21WAF(1)/CIP(1) is critical for the determination of cell fate. In: Cancer Science. 2009 ; Vol. 100, No. 7. pp. 1275-1283.
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