title: [Development of a novel screening assay for inhibitors targeting HIF-1alpha and P300 interaction].
abstract: Hypoxia is a general characteristic of most solid malignancies and intimately related to cancer progression. Homeostatic response to hypoxia is primarily mediated by hypoxia inducible factor-1alpha (HIF-1alpha) that elicits transcriptional activity through recruitment P300 coactivator. Targeting the interaction of HIF- alpha and P300 would thus constitute a novel approach for cancer treatment by suppressing tumor angiogenesis and metastasis. Here, a screening assay was developed for inhibitors targeting the interaction between HIF-1alpha and P300. The nucleotide sequence of human HIF-1alpha and P300 were cloned into pBIND and pACT vectors, named pBIND-HIF1alpha and pACT-P300. The interaction of HIF-1alpha and P300 was identified in HEK293 cell using mammalian two-hybrid system. And compound chetomin decreased their interaction in this mammalian two-hybrid system. We further verified HIF-1 inhibition effect of chetomin in U251-HRE cells. Therefore, we established a screening assay combined HIF-1alpha and P300 mammalian two-hybrid system and U251-HRE reporter assay for HIF-1 selective inhibitors.
authors.forename: Fang-Fang
authors.initials: FF
authors.lastname: Lai
authors.forename: Fei
authors.initials: F
authors.lastname: Niu
authors.forename: Han-Ze
authors.initials: HZ
authors.lastname: Yang
authors.forename: Wan-Qi
authors.initials: WQ
authors.lastname: Zhou
authors.forename: Xiao-Guang
authors.initials: XG
authors.lastname: Chen
mesh_terms: D015687:Cell Hypoxia; D004220:Disulfides / Q000494:pharmacology; D004354:Drug Screening Assays, Antitumor; D050881:E1A-Associated p300 Protein / Q000037:antagonists & inhibitors*; D057809:HEK293 Cells; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000037:antagonists & inhibitors*; D026121:Indole Alkaloids / Q000494:pharmacology; D020798:Two-Hybrid System Techniques
chemical_list: D004220:Disulfides; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D026121:Indole Alkaloids; C001598:chetomin; D050881:E1A-Associated p300 Protein; C494360:EP300 protein, human