The induction of growth arrest DNA damage-inducible gene 45 β in human hepatoma cell lines by S-adenosylmethionine

Weihua Qiu, Bingsen Zhou, Peiguo G. Chu, Frank Luh, Yun Yen

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Down-regulation of GADD45β, which is known to influence cell growth control, apoptosis, and cellular response to DNA damage, has been verified to be specific in hepatocellular carcinoma and consistent with the degree of malignancy. Here, we identified promoter elements for several transcriptional factors in the proximal promoter of GADD45β using the luciferase assay. As a methyl donor for biological transmethylation reactions, S-adenosylmethlonine (SAMe) could restore GADD45β expression in HepG2 in Northern blot analyses and quantitative real-time polymerase chain reaction. Activity and binding capacity of nuclear factor (NF)-κB were confirmed to be specifically induced by SAMe, as evidenced by electrophoretic mobility shift assay, enzyme-linked immunosorbent assay, and a decrease of IκBα in Western blot analyses. The most upstream NF-κB-binding site was crucial for transcriptional activation. In contrast to NF-κB, although there is an E2F-1-binding site adjacent to the NF-κB sites, treatment with SAMe could not induce E2F-1-binding activity. Despite showing a similar GADD45β promoter regulatory pattern as HepG2 (p53 wild type), Hep3B (p53-null) did not exhibit GADD45β induction by SAMe, and the induction could be partially recovered on reconstituting p53 in Hep3B. Thus, our results suggest that GADD45β induction by SAMe via NF-κB may represent a novel mechanism of SAMe-mediated hepatoprotection, with p53 playing an important role.

Original languageEnglish
Pages (from-to)287-296
Number of pages10
JournalAmerican Journal of Pathology
Volume171
Issue number1
DOIs
Publication statusPublished - Jan 1 2007
Externally publishedYes

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S-Adenosylmethionine
DNA Damage
Hepatocellular Carcinoma
Binding Sites
Cell Line
Electrophoretic Mobility Shift Assay
Growth
Luciferases
Northern Blotting
Transcriptional Activation
Genes
Real-Time Polymerase Chain Reaction
Down-Regulation
Western Blotting
Enzyme-Linked Immunosorbent Assay
Apoptosis
Neoplasms
Therapeutics

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

The induction of growth arrest DNA damage-inducible gene 45 β in human hepatoma cell lines by S-adenosylmethionine. / Qiu, Weihua; Zhou, Bingsen; Chu, Peiguo G.; Luh, Frank; Yen, Yun.

In: American Journal of Pathology, Vol. 171, No. 1, 01.01.2007, p. 287-296.

Research output: Contribution to journalArticle

Qiu, Weihua ; Zhou, Bingsen ; Chu, Peiguo G. ; Luh, Frank ; Yen, Yun. / The induction of growth arrest DNA damage-inducible gene 45 β in human hepatoma cell lines by S-adenosylmethionine. In: American Journal of Pathology. 2007 ; Vol. 171, No. 1. pp. 287-296.
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