BTG3 tumor suppressor gene promoter demethylation, histone modification and cell cycle arrest by genistein in renal cancer

Shahana Majid, Altaf A. Dar, Ardalan E. Ahmad, Hiroshi Hirata, Kazumori Kawakami, Varahram Shahryari, Sharanjot Saini, Yuichiro Tanaka, Angela V. Dahiya, Gaurav Khatri, Rajvir Dahiya

Research output: Contribution to journalArticle

Abstract

BTG3/ANA/APRO4 has been reported to be a tumor suppressor gene in some malignancies. It constitutes important negative regulatory mechanism for Src-mediated signaling, a negative regulator of the cell cycle and inhibits transcription factor E2F1. We report that BTG3 is downregulated in renal cancer and that the mechanism of inactivation is through promoter hypermethylation. Quantitative real-time polymerase chain reaction (PCR) showed that BTG3 was downregulated in cancer tissues and cells. Genistein and 5-aza-2′-deoxycytidine (5Aza-C) induced BTG3 messenger RNA (mRNA) expression in A498, ACHN and HEK-293 renal cell carcinoma (RCC) cell lines. Bisulfite-modified PCR and DNA sequencing results showed complete methylation of BTG3 promoter in tumor samples and cancer cell lines. Genistein and 5Aza-C treatment significantly decreased promoter methylation, reactivating BTG3 expression. Chromatin immunoprecipitation assay revealed that genistein and 5Aza-C increased levels of acetylated histones 3, 4, 2H3K4, 3H3K4 and RNA polymerase II at the BTG3 promoter indicative of active histone modifications. Enzymatic assays showed genistein and 5Aza-C decreased DNA Methyltransferase, methyl-CpG-binding domain 2 activity and increased HAT activity. Cell cycle and 3-(4,5-dimethylthiazole-2-yl)-2,5-biphenyl tetrazolium bromide cell proliferation assays showed that genistein has antiproliferative effect on cancer cell growth through induction of cell cycle arrest. This is the first report to show that BTG3 is epigenetically silenced in RCC and can be reactivated by genistein-induced promoter demethylation and active histone modification. Genistein had similar effects to that of 5Aza-C, which is a potent demethylating agent with high toxicity and instability. Genistein being a natural, non-toxic, dietary isoflavone is effective in retarding the growth of RCC cells, making it a promising candidate for epigenetic therapy in renal carcinoma.

Original languageEnglish (US)
Pages (from-to)662-670
Number of pages9
JournalCarcinogenesis
Volume30
Issue number4
DOIs
StatePublished - Apr 17 2009
Externally publishedYes

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Histone Code
decitabine
Genistein
Kidney Neoplasms
Cell Cycle Checkpoints
Tumor Suppressor Genes
Renal Cell Carcinoma
Methylation
Neoplasms
Cell Cycle
Down-Regulation
E2F1 Transcription Factor
Cell Line
Isoflavones
RNA Polymerase II
Chromatin Immunoprecipitation
HEK293 Cells
Methyltransferases
Enzyme Assays
Growth

ASJC Scopus subject areas

  • Cancer Research

Cite this

Majid, S., Dar, A. A., Ahmad, A. E., Hirata, H., Kawakami, K., Shahryari, V., ... Dahiya, R. (2009). BTG3 tumor suppressor gene promoter demethylation, histone modification and cell cycle arrest by genistein in renal cancer. Carcinogenesis, 30(4), 662-670. https://doi.org/10.1093/carcin/bgp042

BTG3 tumor suppressor gene promoter demethylation, histone modification and cell cycle arrest by genistein in renal cancer. / Majid, Shahana; Dar, Altaf A.; Ahmad, Ardalan E.; Hirata, Hiroshi; Kawakami, Kazumori; Shahryari, Varahram; Saini, Sharanjot; Tanaka, Yuichiro; Dahiya, Angela V.; Khatri, Gaurav; Dahiya, Rajvir.

In: Carcinogenesis, Vol. 30, No. 4, 17.04.2009, p. 662-670.

Research output: Contribution to journalArticle

Majid, S, Dar, AA, Ahmad, AE, Hirata, H, Kawakami, K, Shahryari, V, Saini, S, Tanaka, Y, Dahiya, AV, Khatri, G & Dahiya, R 2009, 'BTG3 tumor suppressor gene promoter demethylation, histone modification and cell cycle arrest by genistein in renal cancer', Carcinogenesis, vol. 30, no. 4, pp. 662-670. https://doi.org/10.1093/carcin/bgp042
Majid, Shahana ; Dar, Altaf A. ; Ahmad, Ardalan E. ; Hirata, Hiroshi ; Kawakami, Kazumori ; Shahryari, Varahram ; Saini, Sharanjot ; Tanaka, Yuichiro ; Dahiya, Angela V. ; Khatri, Gaurav ; Dahiya, Rajvir. / BTG3 tumor suppressor gene promoter demethylation, histone modification and cell cycle arrest by genistein in renal cancer. In: Carcinogenesis. 2009 ; Vol. 30, No. 4. pp. 662-670.
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