Reference - Detail
|Author||Nagashima S, Maruyama J, Kawano S, Iwasa H, Nakagawa K, Ishigami-Yuasa M, Kagechika H, Nishina H, Hata Y.|
|Title||Validation of chemical compound library screening for transcriptional co-activator with PDZ-binding motif inhibitors using GFP-fused transcriptional co-activator with PDZ-binding motif.|
Transcriptional co-activator with PDZ-binding motif (TAZ) plays versatile roles in cell proliferation and differentiation. It is phosphorylated by large tumor suppressor kinases, the core kinases of the tumor-suppressive Hippo pathway. Phosphorylation induces the cytoplasmic accumulation of TAZ and its degradation. In human cancers, the deregulation of the Hippo pathway and gene amplification enhance TAZ activity. TAZ interacts with TEA domain family members (TEAD), and upregulates genes implicated in epithelial-mesenchymal transition. It also confers stemness to cancer cells. Thus, TAZ activation provides cancer cells with malignant properties and worsens the clinical prognosis. Therefore, TAZ attracts attention as a therapeutic target in cancer therapy. We applied 18 606 small chemical compounds to human osteosarcoma U2OS cells expressing GFP-fused TAZ (GFP-TAZ), monitored the subcellular localization of GFP-TAZ, and selected 33 compounds that shifted GFP-TAZ to the cytoplasm. Unexpectedly, only a limited number of compounds suppressed TAZ-mediated enhancement of TEAD-responsive reporter activity. Moreover, the compounds that weakened TEAD reporter activity did not necessarily decrease the unphosphorylated TAZ. In this study, we focused on three compounds that decreased both TEAD reporter activity and unphosphorylated TAZ, and treated several human cancer cells with these compounds. One compound did not show a remarkable effect, whereas the other two compounds compromised the cell viability in certain cancer cells. In conclusion, the GFP-TAZ-based assay can be used as the first screening for compounds that inhibit TAZ and show anticancer properties. To develop anticancer drugs, we need additional assays to select the compounds.
|MeSH||Amino Acid Motifs Cell Line, Tumor Cell Nucleus / drug effects Cell Nucleus / metabolism Cell Survival / drug effects Cytoplasm / drug effects Cytoplasm / metabolism Dobutamine / pharmacology Drug Evaluation, Preclinical / methods Drug Evaluation, Preclinical / standards* Ethanolamines / analysis Ethanolamines / pharmacology Genes, Reporter Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism* HEK293 Cells Heterocyclic Compounds, 3-Ring / analysis Heterocyclic Compounds, 3-Ring / pharmacology Humans PDZ Domains / drug effects* Phosphoric Monoester Hydrolases / metabolism Phosphorylation / drug effects Protein Binding / drug effects Protein-Serine-Threonine Kinases / metabolism Pyridines / analysis Pyridines / pharmacology Recombinant Fusion Proteins / chemistry Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Signal Transduction / drug effects Small Molecule Libraries / analysis* Small Molecule Libraries / pharmacology* Thiourea / analogs & derivatives Thiourea / analysis Thiourea / pharmacology Time Factors Transcription Factors / antagonists & inhibitors* Transcription Factors / chemistry* Transcription Factors / genetics Transcription Factors / metabolism Transcription, Genetic / drug effects* ortho-Aminobenzoates / analysis ortho-Aminobenzoates / pharmacology|
|DNA material||pCIneoFLAG-LATS1 (RDB14396) pCIneoFHF-PP1A (RDB14397) pCIneoFHF-PP2A (RDB14398) pCIneoLuc-TAZ (RDB14399) pCIneoFH-TAZ (RDB14400) pCIneoEGFPC2 (RDB14401) pLL3.7-FLAG-YAP1 (RDB14395)|