Suppression of survivin promoter activity by YM155 involves disruption of Sp1-DNA interaction in the survivin core promoter

Qiuying Cheng, Xiang Ling, Andrew Haller, Takahito Nakahara, Kentaro Yamanaka, Aya Kita, Hiroshi Koutoku, Masahiro Takeuchi, Michael G Brattain, Fengzhi Li Dr.

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

YM155, a novel survivin suppressant, shows potent antitumor activity against various human cancers and is currently in phase II clinical trials. In this study, we investigated whether YM155 selectively inhibits survivin transcription. We hypothesize that inhibition of survivin transcription plays a role in YM155-mediated survivin inhibition. We found that YM155 inhibited survivin promoter activity, while it showed minimal inhibitory effect on four control gene promoters in transfection and luciferase activity assay experiments, indicating its selectivity. Transfection of various survivin promoter-luciferase constructs followed by luciferase assays revealed that the survivin core promoter (269 bp) plays a major role in YM155-mediated inhibitory effects. However, flow cytometry analysis indicated that inhibition of survivin promoter activity by YM155 is cell cycle-independent without G1 cell arrests. Electrophoretic mobility shift assays (EMSA) identified that YM155 abrogates nuclear proteins binding to the region of -149 to -71, in which Sp1 is a major candidate, and that YM155 treatment induces Sp1 re-subcellular localization without inhibiting its expression. Forced expression of Sp1 neutralized YM155-mediated downregulation of survivin promoter activity. Consistently, mutation of the identified Sp1 sites in the oligonucleotide probe diminished DNA-protein interactions in EMSA experiments, and mutation of the Sp1 sites in the survivin promoter-luciferase construct diminished survivin promoter activity. These findings indicate that YM155 inhibition of survivin expression is at least in part through its inhibition of survivin transcription by disruption of Sp1 interaction with the region of -149 to -71 in the survivin core promoter.

Original languageEnglish (US)
Pages (from-to)179-197
Number of pages19
JournalInternational Journal of Biochemistry and Molecular Biology
Volume3
Issue number2
StatePublished - Jun 22 2012

Fingerprint

Luciferases
Assays
Transcription
Electrophoretic mobility
DNA
Electrophoretic Mobility Shift Assay
Transfection
Phase II Clinical Trials
Mutation
Flow cytometry
Oligonucleotide Probes
Nuclear Proteins
Protein Binding
Cell Cycle
Flow Cytometry
Down-Regulation
Genes
Experiments
Cells
Neoplasms

Keywords

  • Cancer cells
  • The survivin promoter
  • YM155

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Suppression of survivin promoter activity by YM155 involves disruption of Sp1-DNA interaction in the survivin core promoter. / Cheng, Qiuying; Ling, Xiang; Haller, Andrew; Nakahara, Takahito; Yamanaka, Kentaro; Kita, Aya; Koutoku, Hiroshi; Takeuchi, Masahiro; Brattain, Michael G; Li Dr., Fengzhi.

In: International Journal of Biochemistry and Molecular Biology, Vol. 3, No. 2, 22.06.2012, p. 179-197.

Research output: Contribution to journalArticle

Cheng, Q, Ling, X, Haller, A, Nakahara, T, Yamanaka, K, Kita, A, Koutoku, H, Takeuchi, M, Brattain, MG & Li Dr., F 2012, 'Suppression of survivin promoter activity by YM155 involves disruption of Sp1-DNA interaction in the survivin core promoter', International Journal of Biochemistry and Molecular Biology, vol. 3, no. 2, pp. 179-197.
Cheng, Qiuying ; Ling, Xiang ; Haller, Andrew ; Nakahara, Takahito ; Yamanaka, Kentaro ; Kita, Aya ; Koutoku, Hiroshi ; Takeuchi, Masahiro ; Brattain, Michael G ; Li Dr., Fengzhi. / Suppression of survivin promoter activity by YM155 involves disruption of Sp1-DNA interaction in the survivin core promoter. In: International Journal of Biochemistry and Molecular Biology. 2012 ; Vol. 3, No. 2. pp. 179-197.
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