RESUMO
Various heart diseases cause cardiac remodeling, which in turn leads to ineffective contraction. Although it is an adaptive response to injury, cardiac fibrosis contributes to this remodeling, for which the reactivation of quiescent myofibroblasts is a key feature. In the present study, we investigated the role of the p300/CBP-associated factor (PCAF), a histone acetyltransferase, in the activation of cardiac fibroblasts. An intraperitoneal (i.p.) injection of a high dose (160 mg/kg) of isoproterenol (ISP) induced cardiac fibrosis and reduced the amount of the PCAF in cardiac fibroblasts in the mouse heart. However, the PCAF activity was significantly increased in cardiac fibroblasts, but not in cardiomyocytes, obtained from ISP-administered mice. An in vitro study using human cardiac fibroblast cells recapitulated the in vivo results; an treatment with transforming growth factor-ß1 (TGF-ß1) reduced the PCAF, whereas it activated the PCAF in the fibroblasts. PCAF siRNA attenuated the TGF-ß1-induced increase in and translocation of fibrosis marker proteins. PCAF siRNA blocked TGF-ß1-mediated gel contraction and cell migration. The PCAF directly interacted with and acetylated mothers against decapentaplegic homolog 2 (SMAD2). PCAF siRNA prevented TGF-ß1-induced phosphorylation and the nuclear localization of SMAD2. These results suggest that the increase in PCAF activity during cardiac fibrosis may participate in SMAD2 acetylation and thereby in its activation.
Assuntos
Fibroblastos/metabolismo , Miocárdio/citologia , Proteína Smad2/metabolismo , Fatores de Transcrição de p300-CBP/metabolismo , Acetilação , Actinas/metabolismo , Animais , Movimento Celular , Núcleo Celular/metabolismo , Colágeno Tipo I/metabolismo , Cadeia alfa 1 do Colágeno Tipo I , Fibrose , Humanos , Isoproterenol , Masculino , Metaloproteinases da Matriz/metabolismo , Camundongos Endogâmicos C57BL , Fosforilação , Transporte Proteico , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Fatores de Transcrição de p300-CBP/genéticaRESUMO
BACKGROUND: Transforming growth factor-ß (TGF-ß) signaling is a double-edged sword in cancer development and progression. TGF-ß signaling plays a tumor suppressive role during the early stages of tumor development but promotes tumor progression in later stages. We have previously identified various mutations of TGF-ß receptor II (TßRII) in human oral squamous cell carcinoma (OSCC) samples. In the present study we analyzed I227T/N236D mutation of TßRII, which was detected in the metastatic lymph node of an OSCC patient. METHODS: The effect of I227T/N236D TßRII mutation on transcriptional activities was measured using DR26 cells, which lack functional TßRII. HSC2 human OSCC cells stably expressing wild-type and I227T/N236D mutant TßRII were generated and used to examine the effect of I227T/N236D TßRII mutation on xenograft tumor growth, in vitro cell proliferation, apoptosis, migration, and invasion. RESULTS: The I227T/N236D mutation of TßRII upregulated TGF-ß signaling and promoted xenograft tumor growth when compared with the wild-type, without affecting the in vitro proliferative capacities. To delineate the differences in proliferative capacities in vivo and in vitro, the apoptotic and survival signals were analyzed following curcumin treatment. Concomitant with apoptotic induction, epidermal growth factor receptor (EGFR) activation was observed upon curcumin treatment, which was further activated in I227T/N236D mutant transfectant cells when compared with wild-type cells. Enhanced EGFR activation correlated with cell survival and apoptotic resistance. Enhanced migratory and invasive capabilities of I227T/N236D mutant cells also depended on EGFR signaling. CONCLUSIONS: These results suggest that enhanced EGFR signaling via upregulated TGF-ß signaling shifted the balance toward survival and promoted cell migration and invasion in I227T/N236D mutant cells, elucidating the role of I227T/N236D mutation of TßRII in OSCC progression.
Assuntos
Carcinoma de Células Escamosas/genética , Neoplasias Bucais/genética , Receptor do Fator de Crescimento Transformador beta Tipo II/metabolismo , Animais , Carcinoma de Células Escamosas/patologia , Humanos , Masculino , Camundongos , Camundongos Nus , Neoplasias Bucais/patologia , Mutação , Fenótipo , Transdução de Sinais , TransfecçãoRESUMO
We demonstrate an electrically tunable binary retarder (ETBR) with a self-aligned liquid crystal (LC) on an anisotropic polymer film produced by photo-assisted imprinting. The ETBR has two parts: a tunable optical layer of an LC and a static optical layer of an imprinted anisotropic polymer film possessing two different in-plane optic axes. The anisotropic polymer film was produced using reactive mesogens spontaneously aligned along the topographic microgrooves by imprinting under the exposure of ultraviolet light. An electrically tunable hybrid wave plate, whose phase retardation varies from a quarter to a half-wave, is constructed using the self-aligned LC layer on the imprinted polymer film that behaves as a quarter wave plate with two alternating optic axes. This approach can be used to design a new class of tunable optical devices with multiple in-plane optic axes.
RESUMO
Heart failure is a leading cause of death and is often accompanied by activation of quiescent cardiac myofibroblasts, which results in cardiac fibrosis. In this study, we aimed to identify novel circular RNAs that regulate cardiac fibrosis. We applied transverse aortic constriction (TAC) for 1, 4, and 8 weeks in mice. RNA sequencing datasets were obtained from cardiac fibroblasts isolated by use of a Langendorff apparatus and then further processed by use of selection criteria such as differential expression and conservation in species. CircSMAD4 was upregulated by TAC in mice or by transforming growth factor (TGF)-ß1 in primarily cultured human cardiac fibroblasts. Delivery of si-circSMAD4 attenuated myofibroblast activation and cardiac fibrosis in mice treated with isoproterenol (ISP). si-circSmad4 significantly reduced cardiac fibrosis and remodeling at 8 weeks. Mechanistically, circSMAD4 acted as a sponge against the microRNA miR-671-5p in a sequence-specific manner. miR-671-5p was downregulated during myofibroblast activation and its mimic form attenuated cardiac fibrosis. miR-671-5p mimic destabilized fibroblast growth factor receptor 2 (FGFR2) mRNA in a sequence-specific manner and interfered with the fibrotic action of FGFR2. The circSMAD4-miR-671-5p-FGFR2 pathway is involved in the differentiation of cardiac myofibroblasts and thereby the development of cardiac fibrosis.
RESUMO
Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is a lipid and protein phosphatase that coordinates various cellular processes. Its activity is regulated by the reversible oxidation of an active-site cysteine residue by H2O2 and thioredoxin. However, the potential role of lipid peroxides in the redox regulation of PTEN remains obscure. To evaluate this, 15-hydroperoxy-eicosatetraenoic acid (15s-HpETE), a lipid peroxide, was employed to investigate its effect on PTEN using molecular and cellular-based assays. Exposure to 15s-HpETE resulted in the oxidation of recombinant PTEN. Reversible oxidation of PTEN was also observed in mouse embryonic fibroblast (MEF) cells treated with a 15s-HpETE and Lipofectamine mixture. The oxidative dimerization of thioredoxin was found simultaneously. In addition, the absence of peroxiredoxin III aggravated 15s-HpETE-induced PTEN oxidation in MEF cells. Our study provides novel insight into the mechanism linking lipid peroxidation to the etiology of tumorigenesis.
Assuntos
Leucotrienos/uso terapêutico , Peróxidos Lipídicos/uso terapêutico , PTEN Fosfo-Hidrolase/efeitos dos fármacos , Peroxirredoxina III/uso terapêutico , Animais , Humanos , Leucotrienos/farmacologia , Peróxidos Lipídicos/farmacologia , Camundongos , Oxirredução , Peroxirredoxina III/farmacologia , TransfecçãoRESUMO
We developed an anisotropic nano-imprinting technique, which combines the exposure of ultraviolet light, to fabricate a patterned optical film of a liquid crystalline polymer (LCP). The nano-imprinted LCP film has two-fold functionality of an in-cell retarder and an alignment layer of liquid crystal (LC) molecules. In addition to the geometrically generated microgrooves, the anisotropic surface interactions of the LCP acquired at a nano-scale level during the imprinting process results in the LC alignment on the patterned LCP film without any surface treatment. The nano-imprinted, quarter wave LCP was implemented into a LC cell which produces half-wave retardation under crossed polarizers.
RESUMO
We demonstrate anisotropic optical films based on liquid crystalline polymer (LCP) using a capillary force lithography (CFL). The fabricated optical films can be used as both an optical component and a self-aligning capability of liquid crystal molecules introduced on the film. Additionally, HA or PA LC can be induced on same material by controlling the water repellency of LCP surface. Moreover, surface anchoring transitions could be controlled by variation of pattern sizes and surface treatment. In this point of view, one thin optical film can act both retarder and alignment layer and then shows good retardation, LC alignment, and transmittance at the same time.