Development of a novel screening assay for inhibitors targeting HIF-1alpha and P300 interaction.
- Author:
Fang-Fang LAI
;
Fei NIU
;
Han-Ze YANG
;
Wan-Qi ZHOU
;
Xiao-Guang CHEN
- Publication Type:Journal Article
- MeSH:
Cell Hypoxia;
Disulfides;
pharmacology;
Drug Screening Assays, Antitumor;
E1A-Associated p300 Protein;
antagonists & inhibitors;
HEK293 Cells;
Humans;
Hypoxia-Inducible Factor 1, alpha Subunit;
antagonists & inhibitors;
Indole Alkaloids;
pharmacology;
Two-Hybrid System Techniques
- From:
Acta Pharmaceutica Sinica
2014;49(6):849-853
- CountryChina
- Language:Chinese
-
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.
