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Molecular Cancer Therapeutics
Molecular Cancer Therapeutics
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Article

Acyclic retinoid activates retinoic acid receptor β and induces transcriptional activation of p21CIP1 in HepG2 human hepatoma cells

Masumi Suzui, Masahito Shimizu, Muneyuki Masuda, Jin T. E. Lim, Naoki Yoshimi and I. Bernard Weinstein
Masumi Suzui
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Masahito Shimizu
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Muneyuki Masuda
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Jin T. E. Lim
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Naoki Yoshimi
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I. Bernard Weinstein
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DOI:  Published March 2004
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Abstract

Acyclic retinoid (ACR), a novel synthetic retinoid, has recently been demonstrated by us to inhibit the in vitro growth of human hepatoma cells, and this effect was associated with decreased expression of cell cycle-related molecules. These results, taken together with previous in vitro and clinical studies with ACR, suggest that this agent may be useful in the chemoprevention and therapy of hepatoma and possibly other human malignancies. In the present study, we further examined the molecular effects of ACR on the HepG2 human hepatoma cell line, focusing on the expression of nuclear retinoid receptors and the cell cycle inhibitor protein p21CIP1. Reverse transcription-PCR assays and Western blot analyses indicated that these cells express retinoic acid receptors (RARs) α, β, and γ, retinoid X receptors (RXRs) α and β, and peroxisome proliferator-activated receptors (PPAR) γ mRNA. Treatment with ACR caused a rapid induction within 3 h of RARβ mRNA and the related protein, but there was no significant change in the levels of the mRNA or proteins for RARs α and γ, RXRs α and β, and PPARγ. There was also a rapid increase in p21CIP1 mRNA and protein in HepG2 cells treated with ACR, and this induction occurred via a p53-independent mechanism. In transient transfection reporter assays, we cotransfected the retinoic acid response element-chloramphenicol acetyltransferase (CAT) reporter gene into HepG2 cells together with a RARβ expression vector. RARβ expression markedly stimulated CAT activity (up to about 4-fold) after the addition of ACR. However, CAT activity in the presence of ACR was only about 2-fold higher than that in the absence of ACR, when cells were cotransfected with RARs α and γ or RXRα. These findings suggest that the growth inhibitory effects of ACR are mediated at least in part through RARβ and that both RARβ and p21CIP1 play critical roles in the molecular mechanisms of growth inhibition induced by ACR.

Footnotes

  • Grant support: Supported by awards from the National Foundation for Cancer Research and the T. J. Martell Foundation (I. B. Weinstein), and a grant from the Ministry of Education, Culture, Sports, Science and Technology of Japan (M. Suzui).

  • The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

    • Accepted December 4, 2003.
    • Received July 21, 2003.
    • Revision received December 2, 2003.
  • American Association for Cancer Research
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Molecular Cancer Therapeutics: 3 (3)
March 2004
Volume 3, Issue 3
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Acyclic retinoid activates retinoic acid receptor β and induces transcriptional activation of p21CIP1 in HepG2 human hepatoma cells
Masumi Suzui, Masahito Shimizu, Muneyuki Masuda, Jin T. E. Lim, Naoki Yoshimi and I. Bernard Weinstein
Mol Cancer Ther March 1 2004 (3) (3) 309-316;

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Acyclic retinoid activates retinoic acid receptor β and induces transcriptional activation of p21CIP1 in HepG2 human hepatoma cells
Masumi Suzui, Masahito Shimizu, Muneyuki Masuda, Jin T. E. Lim, Naoki Yoshimi and I. Bernard Weinstein
Mol Cancer Ther March 1 2004 (3) (3) 309-316;
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Molecular Cancer Therapeutics
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