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1.
Anticancer Res ; 39(11): 6097-6105, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31704837

RESUMO

BACKGROUND/AIM: Colorectal cancer (CRC) is one of the most common in the world and its prevalence is rapidly increasing. Jagged-1-activated Notch signaling by apurinic/apyrimidinic endodeoxyribonuclease 1 (APEX1) promotes CRC, and high expression of Jagged-1 is associated with poor prognosis. However, its clinical implication is unknown. The aim of this study was to investigate the clinical role of Jagged-1-activated Notch signaling by APEX1. MATERIALS AND METHODS: The 5-dimethylthiazol-2-yl) 2, 5-diphenyltetrazolium bromide (MTT) assay was used to evaluate the anti-cancer efficacy of 5-fluorouracil (5-FU), oxaliplatin, and irinotecan. Tissue from CRC patients was analyzed to assess the clinical specificity of Jagged-1 activated by APEX1. RESULTS: The half-maximal inhibitory concentration (IC50) in cells co-expressing APEX1 and Jagged-1 cells was higher than that in cells expressing only APEX1. These results indicated that the simultaneous expression of APEX1 and Jagged-1 might be associated with chemoresistance toward 5-FU, oxaliplatin, and irinotecan. Analysis of tissue from CRC patients revealed that high expression of Jagged-1 was associated with a statistically significantly low response to chemotherapy. CONCLUSION: Overexpression of Jagged-1 by APEX1 might serve as a predictor of response to chemotherapy and of poor prognosis, and moreover may be a therapeutic target for chemotherapy of advanced CRC.


Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias Colorretais/patologia , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/metabolismo , Regulação Neoplásica da Expressão Gênica , Proteína Jagged-1/metabolismo , Receptor Notch1/metabolismo , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Apoptose , Biomarcadores Tumorais/genética , Proliferação de Células , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/metabolismo , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/genética , Feminino , Fluoruracila/administração & dosagem , Seguimentos , Humanos , Irinotecano/administração & dosagem , Proteína Jagged-1/genética , Masculino , Pessoa de Meia-Idade , Oxaliplatina/administração & dosagem , Prognóstico , Receptor Notch1/genética , Estudos Retrospectivos , Taxa de Sobrevida , Células Tumorais Cultivadas
2.
Endocrinology ; 160(12): 2863-2876, 2019 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-31609444

RESUMO

The Notch pathway plays diverse and complex roles in cell signaling during development. In the mammalian ovary, Notch is important for the initial formation and growth of follicles, and for regulating the proliferation and differentiation of follicular granulosa cells during the periovulatory period. This study seeks to determine the contribution of female germ cells toward the initial activation and subsequent maintenance of Notch signaling within somatic granulosa cells of the ovary. To address this issue, transgenic Notch reporter (TNR) mice were crossed with Sohlh1-mCherry (S1CF) transgenic mice to visualize Notch-active cells (EGFP) and germ cells (mCherry) simultaneously in the neonatal ovary. To test the involvement of oocytes in activation of Notch signaling in ovarian somatic cells, we ablated germ cells using busulfan, a chemotherapeutic alkylating agent, or investigated KitWv/Wv (viable dominant white-spotting) mice that lack most germ cells. The data reveal that Notch pathway activation in granulosa cells is significantly suppressed when germ cells are reduced. We further demonstrate that disruption of the gene for the Notch ligand Jag1 in oocytes similarly impacts Notch activation and that recombinant JAG1 enhances Notch target gene expression in granulosa cells. These data are consistent with the hypothesis that germ cells provide a ligand, such as Jag1, that is necessary for activation of Notch signaling in the developing ovary.


Assuntos
Células da Granulosa/metabolismo , Proteína Jagged-1/metabolismo , Oócitos/metabolismo , Receptores Notch/metabolismo , Animais , Linhagem Celular , Feminino , Camundongos , Camundongos Transgênicos
3.
Curr Med Sci ; 39(3): 419-425, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31209813

RESUMO

In order to investigate the role of the Notch signaling pathway in skeletal muscle fibrosis after nerve injury, 60 Sprague-Dawley rats were selected and divided randomly into a control and two experimental groups. Group A served as controls without any treatment. Rats in groups B were injected intraperitoneally with 0.2 mL PBS and those in group C were injected intraperitoneally with 0.2 mL PBS+100 µmol/L, 0.2 mL N-[N-(3,5-difluorophenacetyl)-l-alanyl]- S-phenylglycine t-butyl ester (DAPT, a gamma-secretase inhibitor that suppresses Notch signaling) respectively, on postoperative days 1, 3, 7, 10, and 14 in a model of denervation-induced skeletal muscle fibrosis by right sciatic nerve transection. Five rats from each group were euthanized on postoperative days 1, 7, 14, and 28 to collect the right gastrocnemii, and hematoxylin and eosin (HE) staining, immunohistochemistry test, real-time PCR, and Western blotting were performed to assess connective tissue hyperplasia and fibroblast density as well as expression of Notch 1, Jagged 1, and Notch downstream molecules Hes 1 and collagen I (COL I) on day 28. There was no significant difference in HE-stained fibroblast density between group B and C on postoperative day 1. However, fibroblast density was significantly higher in group B than in group C on postoperative days 7, 14, and 28. Notch 1, Jagged 1, Hes 1, and COL I proteins in the gastrocnemius were expressed at very low levels in group A but at high levels in group B. Expression levels of these proteins were significantly lower in group C than in group B (P<0.05), but they were higher in group C than in group A (P<0.05) on postoperative day 28. We are led to conclude that locking the Notch signaling pathway inhibits fibrosis progression of denervated skeletal muscle. Thus, it may be a new approach for treatment of fibrosis of denervated skeletal muscle.


Assuntos
Secretases da Proteína Precursora do Amiloide/genética , Dipeptídeos/farmacologia , Inibidores Enzimáticos/farmacologia , Fibroblastos/metabolismo , Receptor Notch1/genética , Nervo Isquiático/efeitos dos fármacos , Secretases da Proteína Precursora do Amiloide/antagonistas & inibidores , Secretases da Proteína Precursora do Amiloide/metabolismo , Animais , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Fibroblastos/efeitos dos fármacos , Fibroblastos/patologia , Fibrose/prevenção & controle , Regulação da Expressão Gênica , Músculos Isquiotibiais/efeitos dos fármacos , Músculos Isquiotibiais/inervação , Músculos Isquiotibiais/metabolismo , Injeções Intraperitoneais , Proteína Jagged-1/genética , Proteína Jagged-1/metabolismo , Masculino , Denervação Muscular/métodos , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/inervação , Músculo Esquelético/metabolismo , Ratos , Ratos Sprague-Dawley , Receptor Notch1/antagonistas & inibidores , Receptor Notch1/metabolismo , Nervo Isquiático/lesões , Nervo Isquiático/metabolismo , Transdução de Sinais , Fatores de Transcrição HES-1/genética , Fatores de Transcrição HES-1/metabolismo
4.
Cells ; 8(6)2019 06 06.
Artigo em Inglês | MEDLINE | ID: mdl-31174355

RESUMO

Bone morphogenetic protein (BMP) and Notch signaling are critical for endothelial cell (EC) differentiation in vascular development. Recent studies have shown that excess BMP activity induces Notch signaling in cerebral ECs resulting in arteriovenous malformation (AVMs). However, it is unclear how the crosstalk between BMP and Notch signaling affects cerebral EC differentiation at the gene regulatory level. Here, we report that BMP6 activates the activin receptor-like kinase (ALK) 3, a BMP type 1 receptor, to induce Notch1 receptor and Jagged1 and Jagged2 ligands. We show that increased expression of the Notch components alters the transcriptional regulatory complex in the SRY-Box 2 (Sox2) promoter region so as to induce its expression in cerebral ECs. Together, our results identify Sox2 as a direct target of BMP and Notch signaling and provide information on how altered BMP and Notch signaling affects the endothelial transcriptional landscape.


Assuntos
Proteínas Morfogenéticas Ósseas/metabolismo , Receptor Notch1/metabolismo , Fatores de Transcrição SOXB1/metabolismo , Receptores de Ativinas Tipo I/antagonistas & inibidores , Receptores de Ativinas Tipo I/genética , Receptores de Ativinas Tipo I/metabolismo , Animais , Proteínas Morfogenéticas Ósseas/genética , Encéfalo/citologia , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Expressão Gênica , Humanos , Proteína Jagged-1/genética , Proteína Jagged-1/metabolismo , Proteína Jagged-2/genética , Proteína Jagged-2/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Regiões Promotoras Genéticas , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Receptor Notch1/genética , Fatores de Transcrição SOXB1/genética
5.
Genes Genomics ; 41(8): 919-926, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31041680

RESUMO

BACKGROUND: Ovarian cancer is the one of the most deadly gynecologic malignancy among cancer related death in women. However, the treatment for ovarian cancer is still limited. In this study, we aimed to explore the inhibition potential of miR-377-3p in ovarian cancer and explore the mechanism of this effect. METHODS: Quantitative real-time PCR was used to detect the mRNA or microRNA (miRNA) levels. CCK-8, wound-healing, transwell assay were used to detect cell proliferation, migration and invasion. The protein levels were examined by western blot. The dual luciferase reporter assay was conducted to examine the luciferase activity. Tumor volume was measured and Ki67 was detected via immunohistochemistry. RESULTS: qRT-PCR results showed that miR-377-3p was downregulated in ovarian cancer patients. MiR-377-3p mimics suppressed cell proliferation, migration, invasion and decreased the JAG1 level. However, miR-377-3p inhibitor promoted these appearances. Interestingly, we found JAG1 was a target gene of miR-377-3p. JAG1 overexpression reversed the miR-377-3p-induced inhibition of proliferation and invasion. In addition, miR-377-3p inhibited ovarian cancer tumorigenesis in vivo, indicating by decreased tumor volume and staining of Ki67. CONCLUSION: The results showed that miR-377-3p inhibited growth and invasion of ovarian cancer cells by targeting JAG1.


Assuntos
Proteína Jagged-1/genética , MicroRNAs/genética , Neoplasias Ovarianas/genética , Animais , Linhagem Celular Tumoral , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Proteína Jagged-1/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/metabolismo , Invasividade Neoplásica , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia
6.
J Biotechnol ; 299: 44-49, 2019 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-31039368

RESUMO

Atrial fibrillation (AF) is the most common cardiac arrhythmia affecting 1-2% of the general population. Some common variants located in or next to PITX2 and NEURL1 genes are proved to play role in the occurrence of AF. The aim of our study was to investigate whether rs2595104 in the 4q25 chromosome region and rs6584555 SNP in the NEURL1 gene on chromosome 10 is associated with AF in a Caucasian population. We genotyped DNA samples of 76 AF patients and 77 healthy controls using quantitative real-time PCR followed by melting curve analysis. The minor A allele frequency of rs2595104 in PITX2 was 0.38 and 0.44 in the control group and in AF patients, respectively. There was no significant difference in allele and genotype distribution between the two groups (p = 0.52). The allele frequency based log additive odds ratio is 1.22 (C.I. = 0.76-1.94; p = 0.42). The frequency of minor rs6584555 C allele in NEURL1 was 0.22 in the control group and 0.23 in AF patients. Again there were no significant differences in allele and genotype frequencies between AF patients and controls (p = 0.92). The log additive odds ratio is 1,15 (C.I. = 0.66-2.01; p = 0,63). The heterozygous genotype of rs2595104 had the highest frequency compared to the other genotypes in both groups. In case of the rs6584555 SNP the homozygous genotype of the major allele (TT) had the highest frequency in both groups (0.59). The frequency of homozygous genotype for risk allele had the lowest frequency for both SNPs [rs2595104 (AA): 0.19 in patients, 0.12 in controls; rs6584555 (CC): 0.05 in patients, 0.03 in controls]. We did not find significant association between SNP rs2595104 and rs6584555 andAF. We performed a protein-protein network analysis to assess functional connection among the protein products. The proteins coded by PITX2 and NEURL1 are connected indirectly via CTNNB1 and either JAG1 or DLL4 proteins. These interactive proteins are components of two major channels of cell communication pathways, the Wnt and Notch signaling pathways.


Assuntos
Fibrilação Atrial/genética , Proteínas de Homeodomínio/genética , Polimorfismo de Nucleotídeo Único , Fatores de Transcrição/genética , Ubiquitina-Proteína Ligases/genética , Adulto , Idoso , Fibrilação Atrial/metabolismo , Estudos de Casos e Controles , Grupo com Ancestrais do Continente Europeu/genética , Feminino , Frequência do Gene , Predisposição Genética para Doença , Proteínas de Homeodomínio/metabolismo , Humanos , Hungria , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Proteína Jagged-1/metabolismo , Masculino , Pessoa de Meia-Idade , Mapas de Interação de Proteínas , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Transcrição/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , beta Catenina/metabolismo
7.
Mol Med Rep ; 19(6): 5227-5236, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31059052

RESUMO

Psoriasis is a chronic inflammatory disease characterized by the abnormal differentiation and hyperproliferation of epidermal keratinocytes. The aim of the present study was to investigate the mechanism by which microRNA­125b (miR­125b) inhibits the activation of the bromodomain­containing protein 4 (BRD4)/Notch signaling pathway in psoriasis. The contents of associated miRNAs in serum samples from 32 patients with psoriasis were detected by reverse transcription­quantitative polymerase chain reaction (RT­qPCR). The most significantly downregulated miRNA, miR­125b, was screened out. In experiments using HaCaT cells, the association between miR­125b and cell proliferation was observed using a Cell Counting Kit­8 assay, that between miR­125b and the Notch signaling pathway was observed by western blotting and RT­qPCR, and that between miR­125b and the upstream molecule BRD4 of the Notch signaling pathway was observed by luciferase reporter assay and western blotting. The proliferation of HaCaT cells became apparent following miR­125b inhibition. The Jagged­1 ligand in the Notch signaling pathway was upregulated, the active intracellular domain of the Notch1 receptor was increasingly truncated, and the Notch signaling pathway was activated. Furthermore, the inhibited miR­125b contributed directly toward the upstream protein BRD4 3'­UTR of Jagged­1, ultimately activating the Notch signaling pathway with the upregulation of Jagged­1. In conclusion, the proliferation of HaCaT cells mediated by the Jagged­1/Notch signaling pathway was decreased with the miR­125b­mediated inhibition of BRD4 expression. Therefore, miR­125b may be a biomarker and potential therapeutic target for psoriasis treatment.


Assuntos
Proliferação de Células , Proteína Jagged-1/metabolismo , MicroRNAs/metabolismo , Proteínas Nucleares/metabolismo , Psoríase/patologia , Receptores Notch/metabolismo , Fatores de Transcrição/metabolismo , Regiões 3' não Traduzidas , Adolescente , Adulto , Antagomirs/metabolismo , Proteínas de Ciclo Celular , Linhagem Celular , Sobrevivência Celular , Feminino , Humanos , Masculino , MicroRNAs/antagonistas & inibidores , MicroRNAs/sangue , MicroRNAs/genética , Proteínas Nucleares/antagonistas & inibidores , Proteínas Nucleares/genética , Psoríase/metabolismo , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Transdução de Sinais , Fatores de Transcrição/antagonistas & inibidores , Fatores de Transcrição/genética , Adulto Jovem
8.
Neurol Res ; 41(7): 658-664, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31023175

RESUMO

Objectives: Neurite outgrowth of neurons is essential for forming functional neural circuits. It is believed that neuronal neurite outgrowth is an important mechanism of brain plasticity. Rosuvastatin (RSV) is a relatively new statin and may have neuroprotective properties. However, whether RSV exerts an effect on neurite extension and its potential mechanism in cortical neurons remains poorly documented. Methods: Immunofluorescence method was used to examine the effect of RSV on neurite outgrowth in primary cortical neuron by measuring neurite length and confirmed the promotion effect. Then, the potential mechanisms involving the Notch1 pathway were investigated. Effects of RSV on the expression of Notch 1 and Hes1were determined using qRT-PCR. In addition, brain-derived neurotrophic factor (BDNF) expression was also assessed using qRT-PCR, and ELISA. Results: RSV promoted neurite outgrowth of cortical neurons, and this effect could be partially prevented by the Notch 1 pathway inhibitor, DAPT. Subsequently, we found that Jagged 1 and Notch 1 were colocalized. In addition, we observed that the levels of both Notch 1 and Hes 1 in cortical neurons were increased after RSV, but sharply decreased after DAPT treatment. Moreover, RSV increased brain-derived neurotrophic factor (BDNF) levels in cortical neurons, but in the culture medium, and the effect could be partially suppressed by DAPT treatment. Discussion: These findings indicate that RSV mediates neurite outgrowth in primary cortical neurons. The RSV-induced neuritogenic effect is mediated at least partly via the Notch1/BDNF pathway.


Assuntos
Fator Neurotrófico Derivado do Encéfalo/metabolismo , Córtex Cerebral/citologia , Neuritos/efeitos dos fármacos , Crescimento Neuronal/efeitos dos fármacos , Receptor Notch1/metabolismo , Rosuvastatina Cálcica/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Córtex Cerebral/metabolismo , Dipeptídeos/farmacologia , Proteína Jagged-1/metabolismo , Camundongos , Neuritos/fisiologia , Crescimento Neuronal/fisiologia , Cultura Primária de Células , Receptor Notch1/biossíntese , Rosuvastatina Cálcica/antagonistas & inibidores
9.
Microvasc Res ; 124: 67-75, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30930165

RESUMO

Pathological angiogenesis is a hallmark of many diseases. Previously, we reported that orphan nuclear receptor TR3/Nur77 was a critical mediator of angiogenesis to regulate tumor growth and skin wound healing via regulating the expression of the junctional proteins and integrins. However, the molecular mechanism, by which TR3/Nur77 regulates angiogenesis is not completely understood. Here, we were the first to find that TR3/Nur77, via its various amino acid fragments, regulated the expression of DLL4 and Jagged 1 in cultured endothelial cells. DLL4 and Jagged1 mediated TR3/Nur77-induced angiogenic responses and signaling molecules, but not the expression of integrins. Instead, integrins regulated the expressions of DLL4 and Jagged1 induced by TR3/Nur77. Further, DLL4, Jagged1 and integrins α1, α2, ß3 and ß5 were regulated by TR3/Nur77 in animal sepsis models of lipopolysaccharide (LPS)-induced endotoxemia, and cecal ligation and puncture (CLP), in which, TR3/Nur77 expression was significantly and tranciently increased. Mouse survival rates were greatly increased in Nur77 knockout mice bearing both CLP and LPS models. The results elucidated a novel axis of VEGF/histamine ➔ TR3/Nur77 ➔ integrins ➔ DLL4/Jagged1 in angiogenesis, and demonstrated that TR3/Nur77 was an excellent target for sepsis. These studies supported our previous findings that TR3/Nur77 was an excellent therapeutic target, and further our understanding of the molecular mechanism, by which TR3/Nur77 regulated angiogenesis.


Assuntos
Endotoxemia/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteína Jagged-1/metabolismo , Proteínas de Membrana/metabolismo , Neovascularização Fisiológica , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/metabolismo , Animais , Proteínas de Ligação ao Cálcio , Células Cultivadas , Modelos Animais de Doenças , Endotoxemia/genética , Endotoxemia/patologia , Feminino , Humanos , Integrinas/metabolismo , Masculino , Camundongos Knockout , Neovascularização Patológica , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/deficiência , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Transdução de Sinais
10.
Anticancer Res ; 39(4): 2129-2138, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30952759

RESUMO

BACKGROUND/AIM: Early-stage colorectal cancer (CRC) carries a wide range of survival probabilities. Novel biomarkers in this setting are eagerly awaited. Cancer stem cells (CSCs) are considered one of the reasons for treatment failure. This study sought to determine whether activation of pathways governing the function of CSC's could correlate with treatment outcomes. MATERIALS AND METHODS: Tumor specimens from 325 patients were analyzed with immunohistochemistry (IHC) for Hedgehog and Notch pathway activation and results were correlated with prognosis. RESULTS: Positive Notch3 protein expression was an unfavorable prognostic factor for disease-free survival (DFS) and overall survival (OS) (HR=2.43, p=0.024 and HR=2.56, p=0.028, respectively). Activation of the Shh pathway showed univariately longer DFS (HR=0.49, p=0.032). Possible crosstalk between the two pathways was indicated. No further associations between pathway activation and outcome were evident. CONCLUSION: Apart from Notch 3, activation of the pathways, as indicated by IHC expression of their components, did not result in differences in terms of DFS or OS.


Assuntos
Neoplasias Colorretais/metabolismo , Proteínas Hedgehog/metabolismo , Receptor Notch3/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Intervalo Livre de Doença , Feminino , Humanos , Proteína Jagged-1/metabolismo , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Prognóstico , Receptor Notch2/metabolismo , Transdução de Sinais , Adulto Jovem
11.
Mol Carcinog ; 58(7): 1105-1117, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30828884

RESUMO

The present study aimed to determine whether grape seed extract (GSE) procyanidin mix, and its active constituent procyanidin B2 3,3″-di-O-gallate (B2G2) have the potential to target cancer stem cells (CSCs) in prostate cancer (PCa). The CSC populations were isolated and purified based on CD44+ -α2ß1high surface markers in PCa cell lines LNCaP, C4-2B, 22Rv1, PC3, and DU145, and then subjected to prostasphere formation assays in the absence or presence of GSE or B2G2. Results indicated that at lower doses (<15 µg) , the GSE procyanidin mix produced activity in unsorted prostate cancer antigen (PCA) cells, but not in sorted; however, multiple treatments with low dose GSE over a course of time inhibited sphere formation by sorted PCA CSCs. Importantly, B2G2 demonstrated significant potential to target both unsorted and sorted CSCs at lower doses. As formation of spheroids, under specific in vitro conditions, is a measure of stemness, these results indicated the potential of both GSE and B2G2 to target the self-renewal of CSC in PCa cell lines, though B2G2 was more potent in its efficacy. Subsequent mechanistic studies revealed that both GSE procyanidins and B2G2 strongly decreased the constitutive as well as Jagged1 (Notch1 ligand)-induced activated Notch1 pathway. In totality, these in vitro studies warrant extensive dose-profiling-based assessments in vivo settings to conclusively determine the impact on CSC pool kinetics on the efficacy of both GSE and B2G2 to target PCa growth as well as tumor relapse.


Assuntos
Antocianinas/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Biflavonoides/farmacologia , Catequina/farmacologia , Extrato de Sementes de Uva/farmacologia , Células-Tronco Neoplásicas/efeitos dos fármacos , Proantocianidinas/farmacologia , Neoplasias da Próstata/tratamento farmacológico , Apoptose/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Humanos , Proteína Jagged-1/metabolismo , Masculino , Células-Tronco Neoplásicas/patologia , Células PC-3 , Próstata/patologia , Neoplasias da Próstata/patologia , Receptor Notch1/metabolismo , Esferoides Celulares/efeitos dos fármacos , Células Tumorais Cultivadas
12.
Mol Med Rep ; 19(5): 3642-3648, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30896794

RESUMO

Retinoblastoma is an intraocular malignant tumor that may severely affect vision and represents a life­threatening disease in children. Arctigenin (ATG) is an active compound that exhibits numerous pharmacological activities, which is isolated from the seeds of greater burdock (Arctium lappa Linnaeus), a plant used in traditional Chinese herbal medicine. The present study aimed to investigate the effects of ATG on cancer progression by analyzing the retinoblastoma cell line Y79. ATG exhibited a significant inhibitory effect on the viability of Y79 cells in a dose­dependent manner. Furthermore, treatment with ATG promoted apoptosis, and increased the protein expression levels of B­cell lymphoma 2 (BCL­2)­associated X protein and decreased the protein expression levels of BCL­2. Cell migration was suppressed following treatment with ATG, as assessed by Transwell migration assay. Furthermore, the protein expression levels of jagged­1 (JAG1) were decreased, and various factors involved in the Notch signaling pathway, including the Notch intracellular domain (NICD), transcription factor HES (HES)5 and HES1 were downregulated following treatment with ATG. The decreased expression levels of JAG1 were restored in response to JAG1 overexpression, alongside increases in the protein expression levels of NICD, HES5 and HES1. Furthermore, overexpression of JAG1 partly restored the cell viability and migration suppressed following treatment with ATG. In addition, ATG­induced apoptosis was reduced by JAG1 overexpression. Collectively, the present results suggested that ATG may serve as an antitumor compound by suppressing the proliferation and migration of retinoblastoma cells, inducing apoptosis, downregulating the protein expression levels of JAG1, and decreasing the activity of the Notch signaling pathway.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Furanos/farmacologia , Proteína Jagged-1/metabolismo , Lignanas/farmacologia , Retinoblastoma/metabolismo , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células , Sobrevivência Celular/efeitos dos fármacos , Humanos , Proteína Jagged-1/genética , Receptores Notch/metabolismo , Retinoblastoma/genética , Transdução de Sinais/efeitos dos fármacos
13.
J Med Microbiol ; 68(4): 649-656, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30843783

RESUMO

BACKGROUND: Respiratory syncytial virus (RSV) infection is associated with serious lung disease in infants and immunocompromised individuals and is linked to development of asthma. Infection of RSV has been shown to induce Th lymphocyte differentiation. The present study was designed to determine the effects of RSV on the expression of Notch-1 and the related mechanisms on subsequent differentiation of Th lymphocytes. METHODS: A RSV-infected animal model was established and investigated at 7, 28 and 60 days post infection. Real-time qPCR and Western blot were used to observe the expression levels of Notch-1 in CD4+ T cells and its five ligands in lung tissues. The methylation levels of CpG islands in autoimmune regulator (AIRE) and Notch-1 promoters were analysed by time-of-flight mass spectrometry. The differentiation of Th lymphocytes was assayed by real-time qPCR. The distribution of JAG1 and DLL3 in the lung tissues were assayed by immunohistochemistry. The correlation between Th17 and DLL3 was analysed by simple correlation. RESULTS: The results showed that RSV promoted the expression and de-methylation of Notch-1 promoters in CD4+ T cells. Moreover, RSV infection promoted Th1 differentiation at day 7 and day 28; Th17 differentiation at day 7, day 28 and day 60; Th2 differentiation at day 28 and day 60. At the same time, RSV infection promoted the expression of JAG1 and DLL3. Activation of Notch-1/ DLL 3 in lungs may be associated with the differentiation of Th17 lymphocytes. CONCLUSIONS: Our data showed that activation of RSV stimulated the differentiation of Th17 in airway microenvironment through activation Notch-1/DLL3, which may be associated with the occurrence and development of RSV-induced asthma.


Assuntos
Diferenciação Celular , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Pulmão/imunologia , Proteínas de Membrana/metabolismo , Receptor Notch1/metabolismo , Vírus Sincicial Respiratório Humano/imunologia , Células Th17/citologia , Animais , Líquido da Lavagem Broncoalveolar/imunologia , Líquido da Lavagem Broncoalveolar/virologia , Linfócitos T CD4-Positivos , Microambiente Celular/imunologia , Modelos Animais de Doenças , Peptídeos e Proteínas de Sinalização Intracelular/genética , Proteína Jagged-1/genética , Proteína Jagged-1/metabolismo , Pulmão/virologia , Masculino , Proteínas de Membrana/genética , Metilação , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Reação em Cadeia da Polimerase em Tempo Real , Receptor Notch1/genética , Infecções por Vírus Respiratório Sincicial/imunologia , Células Th1/imunologia , Células Th17/imunologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
14.
Neurobiol Aging ; 76: 80-95, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30708185

RESUMO

Olfaction declines with aging and appears to be a prodromal sign of cognitive decline in progressive neurodegenerative diseases. Nevertheless, very little is known about the pathophysiological changes underlying smell loss that may reflect early network dysfunction. A cross-sectional histoanatomical study was conducted on postmortem olfactory nerves of patients with increasing severity of dementia from mild cognitive impairment (MCI) to moderate and severe Alzheimer's disease. The olfactory bulbs and tracts show a prominent and progressive tauopathy in contrast to a weaker amyloid pathology localized to the glomerular region. Topological analysis of Notch signaling components reveals a transient increase in Jagged1 expression in mitral cells of the olfactory bulb of patients with MCI and a gradual decline onwards. Analysis of the olfactory tract reveals an abundance of corpora amylacea, which declines starting from the MCI stage. With the increasing severity of dementia, corpora amylacea are characterized by a gradual shift in cytoskeletal proteins, tau, MAP2 and glial fibrillary acid protein, as well as by a decrease in their Reelin and Jagged1 content. Our research indicates that the olfactory nerve undergoes early and sequential morphological and signaling alterations that correlate with the development of dementia suggesting that this structure may capture and propagate neuronal network imbalances to connected higher brain centers of the entorhinal cortex and hippocampus.


Assuntos
Doença de Alzheimer/patologia , Bulbo Olfatório/patologia , Transdução de Sinais/fisiologia , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/metabolismo , Doença de Alzheimer/fisiopatologia , Moléculas de Adesão Celular Neuronais/genética , Moléculas de Adesão Celular Neuronais/metabolismo , Estudos Transversais , Progressão da Doença , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Feminino , Expressão Gênica , Humanos , Proteína Jagged-1/metabolismo , Masculino , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Bulbo Olfatório/metabolismo , Bulbo Olfatório/fisiopatologia , Serina Endopeptidases/genética , Serina Endopeptidases/metabolismo , Proteínas tau/genética , Proteínas tau/metabolismo
15.
Oral Dis ; 25(4): 1203-1213, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30776172

RESUMO

OBJECTIVE: Jagged1 regulates several biological functions in human periodontal ligament cells (hPDLs). The present study aimed to evaluate mRNA expression profiling of Jagged1-treated hPDLs using microarray technique. METHODS: Notch ligands, Jagged1, were indirectly immobilized on tissue culture surface. Subsequently, hPDLs were seeded on Jagged1 immobilized surface and maintained in growth medium for 48 hr. Total RNA was collected and processed. Gene expression profiling was examined using microarray technique. Real-time polymerase chain reaction and immunofluorescence staining were employed to determine mRNA and protein expression levels, respectively. Cell proliferation and colony-forming unit assay were performed. Cell cycle was evaluated using propidium iodide staining and flow cytometry analysis. RESULTS: The isolated cells demonstrated fibroblast-like morphology and exhibited the co-expression of CD44, CD90, and CD105 surface markers. After stimulated with Jagged1, the total of 411 genes was differentially expressed, consisting both coding and non-coding genes. For coding genes, 165 and 160 coding genes were upregulated and downregulated, respectively. Pathway analysis revealed that the upregulated genes were mainly involved in cellular interactions, signal transduction, and collagen formation and degradation while the downregulated genes were in the events and phases in cell cycle. Jagged1 significantly decreased cell proliferation, reduced colony-forming unit ability, and induced G0/G1 cell cycle arrest in hPDLs. CONCLUSION: Jagged1 regulates various biological pathways in hPDLs. This gene expression profiling could help to understand the mechanisms potentially involved in the Notch signaling regulation in periodontal homeostasis.


Assuntos
Perfilação da Expressão Gênica , Proteína Jagged-1/genética , Ligamento Periodontal , Transdução de Sinais , Células Cultivadas , Humanos , Proteína Jagged-1/metabolismo , Análise em Microsséries , RNA Mensageiro , Reação em Cadeia da Polimerase em Tempo Real
16.
Arch Oral Biol ; 99: 134-140, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30682716

RESUMO

OBJECTIVES: The present study aimed to investigate the expression of Notch signaling components during osteogenic differentiation in vitro and bone healing in vivo. In addition, the influence of Notch signaling on osteogenic differentiation of human bone-derived cells was examined. METHODS: Gene expression profiling of osteogenic differentiation of human bone marrow-derived mesenchymal stromal cells in vitro (GSE80614) and bone healing period of murine tibial fracture in vivo (GSE99388) was downloaded from Gene Expression Omnibus database. The expression of Notch signaling components was obtained from bioinformatic tools. Human bone-derived cells were isolated from alveolar and iliac bone. Cells were seeded on Jagged1 immobilized surface. Osteogenic marker gene expression and mineralization were examined using real-time polymerase chain reaction and alizarin red s staining, respectively. RESULTS: From bioinformatic analysis of gene expression profiling, various Notch signaling components were differentially expressed during osteogenic differentiation of human bone marrow-derived mesenchymal stromal cells in vitro and bone healing period of murine tibial fracture in vivo. The common genes differentially regulated of these two datasets were Hes1, Aph1a, Nsctn, Furin, Adam17, Hey1, Pcsk5, Nedd4, Jag1, Heyl, Notch3, Dlk1, and Hey2. For an in vitro analysis, the mineral deposition markedly increased after seeding human bone-derived cells on Jagged1 immobilized surface, correspondingly with the increase of ALP mRNA expression. Jagged1 treatment downregulated TWIST2 mRNA expression in both human alveolar and iliac bone-derived cells. CONCLUSION: Notch signaling is regulated during osteogenic differentiation and bone healing. In addition, the activation of Notch signaling promotes osteogenic differentiation in human alveolar and iliac bone-derived cells. Therefore, Notch signaling manipulation could be a useful approach for enhancing bone regeneration.


Assuntos
Calcificação Fisiológica/fisiologia , Proteína Jagged-1/metabolismo , Osteócitos/metabolismo , Osteogênese/fisiologia , Receptores Notch/metabolismo , Transdução de Sinais , Proteína ADAM17/genética , Células Epiteliais Alveolares/efeitos dos fármacos , Células Epiteliais Alveolares/metabolismo , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Regeneração Óssea/efeitos dos fármacos , Regeneração Óssea/fisiologia , Calcificação Fisiológica/efeitos dos fármacos , Proteínas de Ligação ao Cálcio , Proteínas de Ciclo Celular/genética , Diferenciação Celular , Endopeptidases/genética , Furina/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Humanos , Ílio/efeitos dos fármacos , Ílio/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/genética , Proteína Jagged-1/genética , Proteína Jagged-1/farmacologia , Proteínas de Membrana , Células-Tronco Mesenquimais , Camundongos , Ubiquitina-Proteína Ligases Nedd4/genética , Osteócitos/efeitos dos fármacos , Osteogênese/genética , Pró-Proteína Convertase 5 , RNA Mensageiro , Receptor Notch3/genética , Receptores Notch/genética , Proteínas Repressoras/genética , Fraturas da Tíbia/genética , Fraturas da Tíbia/metabolismo , Fatores de Transcrição HES-1/genética
17.
Immunol Lett ; 207: 6-16, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30629982

RESUMO

The present study evaluates role of Notch1 signaling in the regulation of T cell immunity in leprosy. Peripheral blood mononuclear cells from leprosy patients and healthy controls were activated with Mycobacterium leprae antigens along with activation of Notch1 signaling pathway and then lymphoproliferation was analyzed by lymphocytes transformation test and the expression of Notch1 and its ligands DLL1, Jagged1 and Jagged 2, T cell activation marker and Th1-Th2 cytokines on Th cells in PBMCs of study subjects were analyzed by flow cytometry. Further, these parameters were also analyzed after inhibition of Notch1 signaling pathway. Higher percentage of Notch1expressing Th cells were noted in TT/BT cases and higher percentage of DLL1 expressing Th cells in TT/BT and BL/LL cases. M. leprae antigens were found to induce the expression of Jagged1 on Th cells. Interestingly activation of Notch1 signaling pathway induced lymphoproliferation in BL/LL cases in response of PGL-1. Activation of Notch1 signaling was also found to induce the expression of T cell activation markers CD25, CD69 and Th1 cytokine IFN-γ in response to M. leprae antigens. Immunomodulation through Notch1 signaling seen in our study could be helpful in augmenting Th1 response in leprosy.


Assuntos
Antígenos de Bactérias/imunologia , Glicolipídeos/imunologia , Hanseníase/imunologia , Mycobacterium leprae/imunologia , Receptor Notch1/metabolismo , Células Th1/imunologia , Células Th2/imunologia , Antígenos CD/metabolismo , Antígenos de Diferenciação de Linfócitos T/metabolismo , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Progressão da Doença , Citometria de Fluxo , Humanos , Imunomodulação , Interferon gama/metabolismo , Proteína Jagged-1/genética , Proteína Jagged-1/metabolismo , Lectinas Tipo C/metabolismo , Ativação Linfocitária , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Transdução de Sinais , Equilíbrio Th1-Th2
18.
Nephrology (Carlton) ; 24(4): 472-480, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29717517

RESUMO

AIM: To understand the mechanism of long non-coding RNA (LncRNA) HOTAIR on renal interstitial fibrosis (RIF) by regulating Notch1 pathway via the modulation of miR-124. METHODS: Unilateral ureteral occlusion (UUO) was used to construct the RIF rat model. HK-2 cells induced by TGF-ß1 were used for the in vitro experiment, which were divided into five groups: Vehicle, TGF-ß1, si-HOTAIR+TGF-ß1, miR-124 inhibitor+TGF-ß1, and si-HOTAIR+miR-124 inhibitor+TGF-ß1 groups. Quantitative real-time PCR (qRT-PCR) and Western blot were performed to detect the expression of HOTAIR, miR-124, Notch1- and epithelial-to-mesenchymal transition (EMT)-related proteins. RESULTS: Significant elevated HOTAIR and reduced miR-124 were presented in UUO rats and TGF-ß1-induced HK-2 cells in a time-dependent manner, with the increased Jagged1 (JAG1), Notch1, NICD, α-SMA and FN, as well as the decreased E-cadherin (all P < 0.05). Compared with the TGF-ß1 group, cells in the si-HOTAIR+TGF-ß1 group were remarkably declined in cell proliferation and the protein expressions of JAG1, Notch1, NICD, α-SMA, and FN, but dramatically higher in E-cadherin expression (all P < 0.05). However, in comparison with the si-HOTAIR+TGF-ß1 group, cells in the si-HOTAIR+miR-124 inhibitor+TGF-ß1 group were apparently improved in proliferation and the protein expression of JAG1, Notch1, NICD, α-SMA, and FN, but substantially reduced in the level of E-cadherin protein (all P < 0.05). CONCLUSION: Silencing lncRNA HOTAIR can up-regulate miR-124 to block Notch1 pathway, and thereby alleviating EMT and RIF, indicating HOTAIR as a potential target for RIF treatment.


Assuntos
Nefropatias/metabolismo , Túbulos Renais Proximais/metabolismo , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , Receptor Notch1/metabolismo , Animais , Linhagem Celular , Modelos Animais de Doenças , Transição Epitelial-Mesenquimal , Fibrose , Regulação da Expressão Gênica , Humanos , Proteína Jagged-1/genética , Proteína Jagged-1/metabolismo , Nefropatias/genética , Nefropatias/patologia , Nefropatias/prevenção & controle , Túbulos Renais Proximais/efeitos dos fármacos , Túbulos Renais Proximais/patologia , Masculino , MicroRNAs/genética , RNA Longo não Codificante/genética , Ratos Sprague-Dawley , Receptor Notch1/genética , Transdução de Sinais , Fator de Crescimento Transformador beta1/farmacologia , Obstrução Ureteral/genética , Obstrução Ureteral/metabolismo , Obstrução Ureteral/patologia
19.
Lab Invest ; 99(3): 290-304, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-29795127

RESUMO

Atherosclerosis is the most common cause of heart disease and stroke. The use of animal models has advanced our understanding of the molecular signaling that contributes to atherosclerosis. Further understanding of this degenerative process in humans will require human tissue. Plaque removed during endarterectomy procedures to relieve arterial obstructions is usually discarded, but can be an important source of diseased cells. Resected tissue from carotid and femoral endarterectomy procedures were compared with carotid arteries from donors with no known cardiovascular disease. Vascular smooth muscle cells (SMC) contribute to plaque formation and may determine susceptibility to rupture. Notch signaling is implicated in the progression of atherosclerosis, and plays a receptor-specific regulatory role in SMC. We defined protein localization of Notch2 and Notch3 within medial and plaque SMC using immunostaining, and compared Notch2 and Notch3 levels in total plaques with whole normal arteries using immunoblot. We successfully derived SMC populations from multiple endarterectomy specimens for molecular analysis. To better define the protein signature of diseased SMC, we utilized sequential window acquisition of all theoretical spectra (SWATH) proteomic analysis to compare normal carotid artery SMC with endarterectomy-derived SMC. Similarities in protein profile and differentiation markers validated the SMC identity of our explants. We identified a subset of differentially expressed proteins that are candidates as functional markers of diseased SMC. To understand how Notch signaling may affect diseased SMC, we performed Jagged1 stimulation of primary cultures. In populations that displayed significant growth, Jagged1 signaling through Notch2 suppressed proliferation; cultures with low growth potential were non-responsive to Jagged1. In addition, Jagged1 did not promote contractile smooth muscle actin nor have a significant effect on the mature differentiated phenotype. Thus, SMC derived from atherosclerotic lesions show distinct proteomic profiles and have altered Notch signaling in response to Jagged1 as a differentiation stimulus, compared with normal SMC.


Assuntos
Aterosclerose/metabolismo , Aterosclerose/patologia , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Receptores Notch/metabolismo , Idoso , Doenças das Artérias Carótidas/metabolismo , Doenças das Artérias Carótidas/patologia , Proliferação de Células , Células Cultivadas , Endarterectomia , Feminino , Humanos , Imuno-Histoquímica , Proteína Jagged-1/metabolismo , Masculino , Pessoa de Meia-Idade , Placa Aterosclerótica/metabolismo , Placa Aterosclerótica/patologia , Receptor Notch2/metabolismo , Receptor Notch3/metabolismo , Transdução de Sinais
20.
J Mol Neurosci ; 68(3): 377-388, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30094580

RESUMO

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide with neuroprotective and neurotrophic effects. This suggests its influence on the development of teeth, which are, similarly to the nervous system, ectoderm and neural crest derivatives. Our earlier studies have shown morphological differences between wild-type (WT) and PACAP-deficient mice, with upregulated sonic hedgehog (SHH) signaling in the lack of PACAP. Notch signaling is a key element of proper tooth development by regulating apoptosis and cell proliferation. In this study, our main goal was to evaluate the possible effects of PACAP on Notch signaling pathway. Immunohistochemical staining was performed of Notch receptors (Notch1, 2, 3, 4), their ligands [delta-like protein (DLL)1, 3, 4, Jagged1, 2], and intracellular target molecules [CSL (CBF1 humans/Su (H) Drosophila/LAG1 Caenorhabditis elegans transcription factor); TACE (TNF-α converting enzyme), NUMB] in molar teeth of 5-day-old WT, and homozygous and heterozygous PACAP-deficient mice. We measured immunopositivity in the enamel-producing ameloblasts and dentin-producing odontoblasts. Notch2 receptor and DLL1 expression were elevated in ameloblasts of PACAP-deficient mice compared to those in WT ones. The expression of CSL showed similar results both in the ameloblasts and odontoblasts. Jagged1 ligand expression was elevated in the odontoblasts of homozygous PACAP-deficient mice compared to WT mice. Other Notch pathway elements did not show significant differences between the genotype groups. The lack of PACAP leads to upregulation of Notch pathway elements in the odontoblast and ameloblast cells. The underlying molecular mechanisms are yet to be elucidated; however, we propose SHH-dependent and independent processes. We hypothesize that this compensatory upregulation of Notch signaling by the lack of PACAP could represent a salvage pathway in PACAP-deficient animals.


Assuntos
Dente Molar/metabolismo , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/genética , Receptor Notch1/metabolismo , Transdução de Sinais , Ameloblastos/metabolismo , Animais , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Proteína Jagged-1/genética , Proteína Jagged-1/metabolismo , Camundongos , Dente Molar/citologia , Dente Molar/crescimento & desenvolvimento , Odontoblastos/metabolismo , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/deficiência , Receptor Notch1/genética , Regulação para Cima
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