RESUMO
In the present study, we designed a logic circuit upon angiogenic response controlled by a combination of enzyme-functionalized magnetic microparticles operating in human umbilical vein endothelial cells (HUVECs). The reported results represent the first example of a biocomputing system operating in controlling angiogenesis.
Assuntos
Compostos Férricos/metabolismo , Neovascularização Fisiológica/efeitos dos fármacos , Algoritmos , Catalase/metabolismo , Enzimas/metabolismo , Compostos Férricos/química , Glucose Oxidase/metabolismo , Células Endoteliais da Veia Umbilical Humana , Humanos , Peróxido de Hidrogênio/farmacologia , Magnetismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
The natural product withaferin A (WFA) exhibits antitumor and antiangiogenesis activity in vivo, which results from this drug's potent growth inhibitory activities. Here, we show that WFA binds to the intermediate filament (IF) protein, vimentin, by covalently modifying its cysteine residue, which is present in the highly conserved alpha-helical coiled coil 2B domain. WFA induces vimentin filaments to aggregate in vitro, an activity manifested in vivo as punctate cytoplasmic aggregates that colocalize vimentin and F-actin. WFA's potent dominant-negative effect on F-actin requires vimentin expression and induces apoptosis. Finally, we show that WFA-induced inhibition of capillary growth in a mouse model of corneal neovascularization is compromised in vimentin-deficient mice. These findings identify WFA as a chemical genetic probe of IF functions, and illuminate a potential molecular target for withanolide-based therapeutics for treating angioproliferative and malignant diseases.
Assuntos
Antineoplásicos/farmacologia , Ergosterol/análogos & derivados , Vimentina/metabolismo , Inibidores da Angiogênese/química , Inibidores da Angiogênese/farmacologia , Inibidores da Angiogênese/uso terapêutico , Animais , Antineoplásicos/química , Antineoplásicos/uso terapêutico , Apoptose/efeitos dos fármacos , Sítios de Ligação , Western Blotting , Linhagem Celular , Neovascularização da Córnea/tratamento farmacológico , Eletroforese em Gel Bidimensional , Ergosterol/química , Ergosterol/farmacologia , Ergosterol/uso terapêutico , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Citometria de Fluxo , Humanos , Camundongos , Camundongos Knockout , Modelos Moleculares , Estrutura Molecular , Ligação Proteica , Vimentina/genética , VitanolídeosRESUMO
The major estrogen metabolite 2-methoxyestradiol (2ME) has been shown to target tumor cells without severe side effects and is currently being evaluated in clinical trials for several types of cancer. Despite its promise for use in clinical setting, the mechanism(s) by which 2ME exerts its anti-tumor activity is not clearly defined at this time. Employing organic chemistry tools, we synthesized 2ME analogs with which 2ME affinity column was prepared, enabling us to detect a protein that selectively interacts with 2ME. This 2ME analog will be useful as a probe to identify the biological target(s) of 2ME and study their functions in tumor cells.
