RESUMO
The most active anticancer component in green tea is epigallocatechin-3-gallate (EGCG). The human peptidyl prolyl cis/trans isomerase (Pin1) plays a critical role in oncogenic signaling. Herein, we report the X-ray crystal structure of the Pin1/EGCG complex resolved at 1.9 Å resolution. Notably, the structure revealed the presence of EGCG in both the WW and PPIase domains of Pin1. The direct binding of EGCG with Pin1 was confirmed and the interaction inhibited Pin1 PPIase activity. In addition, proliferation of cells expressing Pin1 was inhibited and tumor growth in a xenograft mouse model was suppressed. The binding of EGCG with Arg17 in the WW domain prevented the binding of c-Jun, a well-known Pin1 substrate. EGCG treatment corresponded with a decreased abundance of cyclin D1 and diminution of 12-O-tetradecanoylphorbol-l3-acetate-induced AP-1 or NF-κB promoter activity in cells expressing Pin1. Overall, these results showed that EGCG directly suppresses the tumor-promoting effect of Pin1.
Assuntos
Catequina/análogos & derivados , Regulação Neoplásica da Expressão Gênica , Peptidilprolil Isomerase/metabolismo , Animais , Catequina/uso terapêutico , Ciclina D1/metabolismo , Glutationa Transferase/metabolismo , Humanos , Camundongos , Camundongos Knockout , Camundongos Nus , NF-kappa B/metabolismo , Peptidilprolil Isomerase de Interação com NIMA , Transplante de Neoplasias , Estrutura Terciária de Proteína , Proteínas Proto-Oncogênicas c-jun/metabolismo , Acetato de Tetradecanoilforbol/metabolismo , Fator de Transcrição AP-1/metabolismoRESUMO
Three new potential hENT(1) inhibitors suitable for labeling with PET/SPECT radioisotopes were prepared from an advanced intermediate 4. They were tested for their capability to inhibit binding of SAENTA-fluorescein to HL60 leukemia cells in flow cytometry assay and SAENTA-I (5) was determined to be the most active compound. (131)I-5 showed high hENT(1)-specific binding (up to 54% ID) to 6 from 7 tested tumor cell lines and was chosen for further in vivo study.
Assuntos
Adenosina/análogos & derivados , Benzamidas/síntese química , Transportador Equilibrativo 1 de Nucleosídeo/metabolismo , Compostos Radiofarmacêuticos/síntese química , Tionucleosídeos/química , Adenosina/síntese química , Adenosina/química , Benzamidas/química , Linhagem Celular Tumoral , Citometria de Fluxo , Corantes Fluorescentes/química , Humanos , Radioisótopos do Iodo/química , Tomografia por Emissão de Pósitrons , Compostos Radiofarmacêuticos/química , Tionucleosídeos/síntese químicaRESUMO
Hydrocortisone, progesterone, testosterone, triiodothyronine, thyroxine, chorionic gonadotropin, prolactin, alpha-fetoprotein, luteinizing, follicle-stimulating, and thyrotropic hormones were measured in human sera and in Lyphochek Immunoassay Plus Control reference sera (Bio-Rad Laboratories, USA) using 4 commercial kits (Alkor Bio Inc. and Roche, automated analyzer Roche Cobas Core; DPC, automated analyzer Immulite; Bayer, automated analyzer ACS:180). Coordination and correlation between these kits was observed, the coordination decreasing in the series Alkor Bio/Bayer, Alkor Bio/Roche, and Alkor Bio/DPC.
