Int J Biochem Mol Biol 2012;3(2):179-197
Original Article
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
Department of Pharmacology & Therapeutics, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, New York, USA; Drug Discovery
Research, Astellas Pharma Inc., Tsukuba, Ibaraki, Japan; Current address: University of Nebraska Medical Center, Omaha, Nebraska, USA
Received March 23, 2012; accepted April 18, 2012; Epub May 18, 2012; Published June 15, 2012
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. (IJBMB1203007)
Keywords: YM155, the survivin promoter, cancer cells
Address all correspondence to:
Dr. Fengzhi Li
Department of Pharmacology and Therapeutics
Roswell Park Cancer Institute, Elm and Carlton Streets
Buffalo, New York 14263, USA.
Tel: (716)-845-4398; Fax: (716)-845-8857
E-mail: fengzhi.li@roswellpark.org
Original Article
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
Department of Pharmacology & Therapeutics, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, New York, USA; Drug Discovery
Research, Astellas Pharma Inc., Tsukuba, Ibaraki, Japan; Current address: University of Nebraska Medical Center, Omaha, Nebraska, USA
Received March 23, 2012; accepted April 18, 2012; Epub May 18, 2012; Published June 15, 2012
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. (IJBMB1203007)
Keywords: YM155, the survivin promoter, cancer cells
Address all correspondence to:
Dr. Fengzhi Li
Department of Pharmacology and Therapeutics
Roswell Park Cancer Institute, Elm and Carlton Streets
Buffalo, New York 14263, USA.
Tel: (716)-845-4398; Fax: (716)-845-8857
E-mail: fengzhi.li@roswellpark.org
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