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1.
Gene ; 717: 144047, 2019 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-31421190

RESUMO

BACKGROUND: Vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF) signaling pathways play important roles in the formation of the blood vascular system and nervous system across animal phyla. We have earlier reported VEGF and FGF from Hydra vulgaris Ind-Pune, a cnidarian with a defined body axis, an organized nervous system and a remarkable ability of regeneration. We have now identified three more components of VEGF and FGF signaling pathways from hydra. These include FGF-1, FGF receptor 1 (FGFR-1) and VEGF receptor 2 (VEGFR-2) with a view to deciphering their possible roles in regeneration. METHODS: In silico analysis of proteins was performed using Clustal omega, Swiss model, MEGA 7.0, etc. Gene expression was studied by whole mount in situ hybridization. VEGF and FGF signaling was inhibited using specific pharmacological inhibitors and their effects on head regeneration were studied. RESULTS: Expression patterns of the genes indicate a possible interaction between FGF-1 and FGFR-1 and also VEGF and VEGFR-2. Upon treatment of decapitated hydra with pharmacological inhibitor of FGFR-1 or VEGFR-2 for 48 h, head regeneration was delayed in treated as compared to untreated, control regenerates. When we studied the expression of head specific genes HyBra1 and HyKs1 and tentacle specific gene HyAlx in control and treated regenerates using whole mount in situ hybridization, expression of all the three genes was found to be adversely affected in treated regenerates. CONCLUSIONS: The results suggest that VEGF and FGF signaling play important roles in regeneration of hypostome and tentacles in hydra.


Assuntos
Fatores de Crescimento de Fibroblastos/metabolismo , Cabeça/fisiologia , Hydra/fisiologia , Regeneração/fisiologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Simulação por Computador , Fator 1 de Crescimento de Fibroblastos/química , Fator 1 de Crescimento de Fibroblastos/genética , Fator 1 de Crescimento de Fibroblastos/metabolismo , Regulação da Expressão Gênica , Humanos , Hydra/efeitos dos fármacos , Indóis/farmacologia , Domínios Proteicos , Pirróis/farmacologia , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/química , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/genética , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/metabolismo , Regeneração/efeitos dos fármacos , Transdução de Sinais , Homologia Estrutural de Proteína , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/química , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genética , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo
2.
Cells ; 8(8)2019 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-31426340

RESUMO

Attention Deficit Hyperactivity Disorder (ADHD) is a highly heritable and prevalent neurodevelopmental disorder that frequently persists into adulthood. Strong evidence from genetic studies indicates that single nucleotide polymorphisms (SNPs) harboured in the ADGRL3 (LPHN3), SNAP25, FGF1, DRD4, and SLC6A2 genes are associated with ADHD. We genotyped 26 SNPs harboured in genes previously reported to be associated with ADHD and evaluated their potential association in 386 individuals belonging to 113 nuclear families from a Caribbean community in Barranquilla, Colombia, using family-based association tests. SNPs rs362990-SNAP25 (T allele; p = 2.46 × 10-4), rs2282794-FGF1 (A allele; p = 1.33 × 10-2), rs2122642-ADGRL3 (C allele, p = 3.5 × 10-2), and ADGRL3 haplotype CCC (markers rs1565902-rs10001410-rs2122642, OR = 1.74, Ppermuted = 0.021) were significantly associated with ADHD. Our results confirm the susceptibility to ADHD conferred by SNAP25, FGF1, and ADGRL3 variants in a community with a significant African American component, and provide evidence supporting the existence of specific patterns of genetic stratification underpinning the susceptibility to ADHD. Knowledge of population genetics is crucial to define risk and predict susceptibility to disease.


Assuntos
Transtorno do Deficit de Atenção com Hiperatividade/genética , Haplótipos/genética , Polimorfismo de Nucleotídeo Único/genética , Adolescente , Adulto , Afro-Americanos/genética , Estudos de Casos e Controles , Criança , Colômbia , Feminino , Fator 1 de Crescimento de Fibroblastos/genética , Predisposição Genética para Doença , Humanos , Masculino , Receptores Acoplados a Proteínas-G/genética , Receptores de Peptídeos/genética , Proteína 25 Associada a Sinaptossoma/genética
3.
Med Sci Monit ; 25: 984-990, 2019 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-30716059

RESUMO

BACKGROUND The expression and mechanism of IL-1, IL-2, IL-8, BMP, FGF1, and IGF-1 in Sprague-Dawley (SD) rats with lumbar disc herniation were investigated. MATERIAL AND METHODS Immunohistochemical methods were applied to identify IL-1, IL-2, IL-8, BMP, FGF1, and IGF-1. PI3K, AKT protein, and mRNA expression were detected and analyzed by Western blot analysis. We selected 30 healthy SD rats and divided them into 2 groups to construct an animal model that was validated by immediate CT scanning. Cartilage tissues from the lumbar disc herniation (experimental) group and control group were obtained and compared. RESULTS The expression of BMP was not significantly different between the control group and the experimental group (P>0.05). FGF1: There was no significant difference in the expression of FGF1 (P>0.05) between the control group and the experimental group. Compared with the control group, the expression of IGF-1 in the experimental group was significantly higher (P<0.05); the expression of IL-1 in the experimental group was significantly higher (P<0.05); and the expression of IL-2 in the experimental group was also significantly higher (P<0.05). There was no significant difference in IL-8 between the experimental group and the control group (P>0.05). The expression levels of PI3K and AKT protein and mRNA were significantly higher than those in healthy controls (P<0.05). CONCLUSIONS After lumbar disc herniation occurred, the IGF-1 was first activated; the PI3K/AKT signaling pathway was later activated, which resulted in the expression of IL-1 and IL-2 inflammation-related factors being increased.


Assuntos
Deslocamento do Disco Intervertebral/metabolismo , Vértebras Lombares/metabolismo , Vértebras Lombares/fisiopatologia , Animais , Proteínas Morfogenéticas Ósseas/genética , Proteínas Morfogenéticas Ósseas/metabolismo , Fator 1 de Crescimento de Fibroblastos/genética , Fator 1 de Crescimento de Fibroblastos/metabolismo , Fator de Crescimento Insulin-Like I/metabolismo , Interleucina-1/genética , Interleucina-1/metabolismo , Interleucina-2/genética , Interleucina-2/metabolismo , Disco Intervertebral , Deslocamento do Disco Intervertebral/fisiopatologia , Região Lombossacral , Masculino , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Ratos , Ratos Sprague-Dawley , Transdução de Sinais
4.
J Cell Mol Med ; 22(12): 6294-6303, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30320493

RESUMO

Diabetic nephropathy (DN) is one of general and common complication of diabetes, which severely affects the physical and mental health of diabetic patients. Fibroblast growth factor 1 (FGF1), an effective control agent of blood glucose, plays an effective treatment role on diabetes-induced renal injury. But the specific molecule mechanism underlying it is still unclear. Since induction of cellular stress is the main and common mechanism of diabetes-induced complication, we hypothesized that reduction of cellular stress is also the molecular mechanism of FGF1 treatment for DN. Here, we have further confirmed that FGF1 significantly ameliorated the diabetes-induced renal interstitial fibrosis and glomerular damage. The expression levels of collagen and α-smooth muscle actin (α-SMA) also dramatically induced in kidney from db/db mice, but these effects were blocked by FGF1 administration. Our mechanistic investigation had further revealed that diabetes significantly induced oxidative stress, nitrosative stress, and endoplasmic reticulum (ER) stress with upregulation of malondialdehyde (MDA), nitrotyrosine level, ER stress makers and downregulation of antioxidant capacity (AOC). FGF1 treatment significantly attenuated the effect of diabetes on cellular stress. We conclude that FGF1-associated glucose decreases and subsequent reduction of cellular stress is the another potential molecule mechanism underlying FGF1 treatment for DN.


Assuntos
Diabetes Mellitus Experimental/genética , Nefropatias Diabéticas/genética , Fator 1 de Crescimento de Fibroblastos/genética , Fibrose/genética , Animais , Antioxidantes/metabolismo , Glicemia/genética , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patologia , Nefropatias Diabéticas/metabolismo , Nefropatias Diabéticas/patologia , Estresse do Retículo Endoplasmático/genética , Fibrose/metabolismo , Fibrose/patologia , Regulação da Expressão Gênica/genética , Humanos , Rim/metabolismo , Rim/patologia , Glomérulos Renais/metabolismo , Glomérulos Renais/patologia , Malondialdeído/metabolismo , Camundongos , Estresse Oxidativo/genética
5.
Biochem Biophys Res Commun ; 505(2): 505-510, 2018 10 28.
Artigo em Inglês | MEDLINE | ID: mdl-30268497

RESUMO

Age-related cataract, the most common cause of blindness worldwide, has been found closely associated with ß-crystallin B2 (ßB2 or CRYBB2). MicroRNAs (miRNAs) are the primary epigenetic regulators important for various biological processes. However, the role of miRNAs in the progression of lens cataract remains to be elucidated. In this study, we found a novel signal cascade miR-326-fibroblast growth factor 1 (FGF1)-ßB2 modulating the progression of lens cataract. In brief, miR-326 exacerbated but its antagomirs attenuated H2O2-induced apoptosis of HLEC-B3 human lens epithelial cells. Dual-luciferase reporter assay and Western blot showed that miR-326 inhibited FGF1 expression by directly targeting its mRNA 3'-UTR. Consistent with this result, miR-326 antagomir enhanced FGF1 protein level. In addition to FGF1, miR-326 antagomir also enhanced ßB2 expression and this enhancement was abolished by transfection of HLEC-B3 cells with FGF1 shRNA. These data demonstrated that miR-326 antagomir increased ßB2 expression via upregulating FGF1, which was further confirmed by the studies in a rat model of selenite-induced cataract. This work suggests that miR-326 antagomir might be a promising candidate to prevent progression of age-related cataract.


Assuntos
Antagomirs/metabolismo , Catarata/metabolismo , Fator 1 de Crescimento de Fibroblastos/metabolismo , MicroRNAs/antagonistas & inibidores , Cadeia B de beta-Cristalina/metabolismo , Regiões 3' não Traduzidas , Fatores Etários , Animais , Apoptose , Catarata/genética , Catarata/patologia , Catarata/terapia , Linhagem Celular , Progressão da Doença , Células Epiteliais/citologia , Fator 1 de Crescimento de Fibroblastos/genética , Humanos , Cristalino/citologia , Ratos Sprague-Dawley , Regulação para Cima
6.
Br Poult Sci ; 59(6): 613-617, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30259763

RESUMO

1. FGF1 and FGF10, two paracrine members of the fibroblast growth factor (FGF) gene family, play critical roles in the development, structural and metabolic remodelling of adipose tissue. 2. The objective of this study was to investigate the expression profiles of FGF1 and FGF10 genes in breast muscle and thigh muscle in 5 developmental stages (1, 81, 119, 154 and 210 d old) in Tibetan chickens. The possible relationships between expression of these genes and intramuscular fat (IMF) content were analysed in Tibetan chickens. 3. Expression profiles showed that FGF1 and FGF10 mRNA were ubiquitously expressed in various tissues of 154-d-old Tibetan chickens. Lung tissue contained the highest amount of FGF1 and FGF10 mRNA while breast muscle and thigh muscle exhibited lower levels of FGF1 and FGF10 mRNA in both males and females compared with other tissues (P < 0.05). Temporal expression of FGF1 and FGF10 in breast and thigh muscle showed similar tendencies in males and females, respectively, with peaks in thigh muscle at 119-d-old and breast muscle in 1-d-old males and females, respectively. 4. Correlation analysis suggested that gender had an influence on the relationships of FGF1 and FGF10 expression with IMF content in thigh muscle. The RNA levels of FGF1 and FGF10 genes in male thigh muscle were positively related to IMF content of Tibetan chickens (P < 0.01), while the correlations were shown to be negative in female thigh muscle (P > 0.05). 5. These results provide a basis for functional elucidation of FGF1 and FGF10 genes on adipocyte development and intramuscular fat deposition, as well as selective breeding and resource exploration of local poultry breeds.


Assuntos
Tecido Adiposo/metabolismo , Galinhas/genética , Fator 10 de Crescimento de Fibroblastos/genética , Fator 1 de Crescimento de Fibroblastos/genética , Músculo Esquelético/metabolismo , Transcriptoma , Tecido Adiposo/química , Tecido Adiposo/crescimento & desenvolvimento , Animais , Feminino , Fator 1 de Crescimento de Fibroblastos/fisiologia , Masculino , Músculo Esquelético/química , RNA Mensageiro/análise , Fatores Sexuais , Tibet
7.
Cancer Lett ; 438: 116-125, 2018 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-30217564

RESUMO

The chromosomal locations of lncRNAs (long non-coding RNAs, lncRNAs) infer their biological functions in cancer. Lnc-RAB1A-2, a Ras-related protein Rab-1A (RAB1A) upstream lncRNA, was chosen for assessment of its impact on lung cancer prognosis in a case-based analysis and investigation of its biological function though a series of functional assays. Lnc-RAB1A-2 was significantly upregulated in 276 lung cancer tissues compared with corresponding non-tumor tissues, and its expression level was significantly correlated with clinical stage and metastasis status in lung cancer patients. Patients with high expression levels of this lncRNA had a shorter median survival time (16.0 months vs. 23.0 months, P = 0.011 in southern samples; 8.0 months vs. 19.0 months, P = 0.020 in eastern samples; 13.0 months vs. 19.0 months, P = 0.002 in merged samples) and a higher risk of death than those with lower levels (HR = 1.52; 95% CI = 1.01-2.26, in merged samples). Additionally, overexpression of lnc-RAB1A-2 significantly promoted lung cancer cell proliferation in vitro and in vivo. Further analyses using digital gene expression tag profiling revealed that lnc-RAB1A-2 could affect the expression of fibroblast growth factor 1 (FGF1), a gene involved in the PI3K/AKT/mTOR pathway that is largely activated by RAB1A. FGF1 was confirmed to be a down-stream gene of lnc-RAB1A-2. Collectively, our study demonstrated that lnc-RAB1A-2 is associated with poor lung cancer prognosis by promoting lung cancer development.


Assuntos
Fator 1 de Crescimento de Fibroblastos/genética , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Neoplasias Pulmonares/genética , RNA Longo não Codificante/genética , Regulação para Cima , Células A549 , Linhagem Celular Tumoral , Proliferação de Células/genética , Feminino , Fator 1 de Crescimento de Fibroblastos/metabolismo , Células HEK293 , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , Transdução de Sinais/genética , Análise de Sobrevida , Transplante Heterólogo , Carga Tumoral/genética
8.
J Mol Biol ; 430(21): 4087-4101, 2018 10 19.
Artigo em Inglês | MEDLINE | ID: mdl-30099027

RESUMO

FGF1 and FGF2 bind to specific cell-surface tyrosine kinase receptors (FGFRs) and activate intracellular signaling that leads to proliferation, migration or differentiation of many cell types. Besides this classical mode of action, under stress conditions, FGF1 and FGF2 are translocated in a receptor-dependent manner via the endosomal membrane into the cytosol and nucleus of the cell. However, despite many years of research, the role of translocated FGF1 and FGF2 inside the cell remains unclear. Here, we reveal an anti-apoptotic activity of intracellular FGF1 and FGF2, which is independent of FGFR activation and downstream signaling. We observed an inhibition of cell apoptosis induced by serum starvation or staurosporine upon treatment with exogenous FGF1 or FGF2, despite the presence of highly potent FGFR inhibitors. Similar results were found when the tyrosine kinase of FGFR1 was completely blocked by a specific mutation. Moreover, the anti-apoptotic effect of the growth factors was abolished by known inhibitors of the translocation of FGF1 and FGF2 from the endosomes to the interior of the cell. Interestingly, FGF2 showed higher anti-apoptotic activity than FGF1. Since FGF2 is not phosphorylated by PKCδ and is present inside the nucleus longer than is FGF1, we speculated that the different activities could reflect their diverse nuclear export kinetics. Indeed, we observed that FGF1 mutations preventing binding to nucleolin and therefore phosphorylation in the nucleus affect the anti-apoptotic activity of FGF1. Taken together, our data indicate that the translocation of FGF1 and FGF2 protects cells against apoptosis and promotes cell survival.


Assuntos
Apoptose , Fator 1 de Crescimento de Fibroblastos/metabolismo , Fator 2 de Crescimento de Fibroblastos/metabolismo , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Linhagem Celular , Expressão Ectópica do Gene , Fator 1 de Crescimento de Fibroblastos/genética , Fator 1 de Crescimento de Fibroblastos/farmacologia , Fator 2 de Crescimento de Fibroblastos/genética , Fator 2 de Crescimento de Fibroblastos/farmacologia , Expressão Gênica , Humanos , Camundongos , Mutação , Inibidores de Proteínas Quinases/farmacologia , Transporte Proteico , Receptores de Fatores de Crescimento de Fibroblastos/antagonistas & inibidores , Receptores de Fatores de Crescimento de Fibroblastos/genética , Receptores de Fatores de Crescimento de Fibroblastos/metabolismo
9.
PLoS One ; 13(7): e0201551, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30063763

RESUMO

MiRNAs play an important role in cell proliferation, apoptosis, and differentiation. MiR-18a is increasingly being recognized as a regulator of cancer pathogenesis. Here, we discovered that miR-18a participates in myoblasts proliferation. Expression of miR-18a was downregulated with the differentiation of C2C12 myoblasts. Overexpression of miR-18a affected the proliferation of C2C12 cells, primary myoblasts and RD cells. MiR-18a influenced the expression of cell cycle-related genes. Using TargetScan 6.2, we found that the 3' untranslated region (UTR) of the mouse Fgf1 gene contains complementary sequences to miR-18a. Using a siRNA, we confirmed that the reduction in the Fgf1 levels inhibited proliferation of C2C12 cells. Therefore, our results show that miR-18a participates in the regulation of proliferation by partly decreasing the expression of Fgf1.


Assuntos
Proliferação de Células/genética , Fator 1 de Crescimento de Fibroblastos/genética , MicroRNAs/fisiologia , Mioblastos/fisiologia , Animais , Células Cultivadas , Regulação da Expressão Gênica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , MicroRNAs/genética
10.
Int J Mol Sci ; 19(9)2018 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-30134556

RESUMO

Fibroblast growth factor 1 (FGF1) and its receptors (FGFRs) regulate crucial biological processes such as cell proliferation and differentiation. Aberrant activation of FGFRs by their ligands can promote tumor growth and angiogenesis in many tumor types, including lung or breast cancer. The development of FGF1-targeting molecules with potential implications for the therapy of FGF1-driven tumors is recently being considered a promising approach in the treatment of cancer. In this study we have used phage display selection to find scFv antibody fragments selectively binding FGF1 and preventing it from binding to its receptor. Three identified scFv clones were expressed and characterized with regard to their binding to FGF1 and ability to interfere with FGF1-induced signaling cascades activation. In the next step the scFvs were cloned to scFv-Fc format, as dimeric Fc fusions prove beneficial in prospective therapeutic application. As expected, scFvs-Fc exhibited significantly increased affinity towards FGF1. We observed strong antiproliferative activity of the scFvs and scFvs-Fc in the in vitro cell models. Presented antibody fragments serve as novel FGF1 inhibitors and can be further utilized as powerful tools to use in the studies on the selective cancer therapy.


Assuntos
Fator 1 de Crescimento de Fibroblastos/antagonistas & inibidores , Biblioteca de Peptídeos , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/antagonistas & inibidores , Proteínas Recombinantes de Fusão/farmacologia , Anticorpos de Cadeia Única/farmacologia , Animais , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Células Clonais , Clonagem Molecular , Reações Cruzadas , Escherichia coli/genética , Escherichia coli/metabolismo , Fator 1 de Crescimento de Fibroblastos/genética , Fator 1 de Crescimento de Fibroblastos/metabolismo , Expressão Gênica , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Humanos , Cinética , Camundongos , Células NIH 3T3 , Ligação Proteica , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/genética , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/metabolismo , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/genética , Transdução de Sinais , Anticorpos de Cadeia Única/biossíntese , Anticorpos de Cadeia Única/genética
11.
J Cell Mol Med ; 22(10): 4760-4770, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30010249

RESUMO

Tumour growth depends on a continual supply of the nutrients and oxygen, which are offered by tumour angiogenesis. Our previous study showed that dipalmitoylphosphatidic acid (DPPA), a bioactive phospholipid, inhibits the growth of triple-negative breast cancer cells. However, its direct effect on angiogenesis remains unknown. Our work showed that DPPA significantly suppressed vascular growth in the chick embryo chorioallantoic membrane (CAM) and yolk sac membrane (YSM) models. Meanwhile, tumour angiogenesis and tumour growth were inhibited by DPPA in the tumour tissues of an experimental breast cancer model, a subcutaneous xenograft mouse model and a genetically engineered spontaneous breast cancer mouse model (MMTV-PyMT). Furthermore, DPPA directly inhibited the proliferation, migration and tube formation of vascular endothelial cells. The anti-angiogenic effect of DPPA was regulated by the inhibition of Cut-like homeobox1 (CUX1), which transcriptionally inhibited fibroblast growth factor 1 (FGF1), leading to the downregulation of hepatocyte growth factor (HGF). This work first demonstrates that DPPA directly inhibits angiogenesis in cancer development. Our previous work along with this study suggest that DPPA functions as an anti-tumour therapeutic drug that inhibits angiogenesis.


Assuntos
Antineoplásicos/farmacologia , Fator 1 de Crescimento de Fibroblastos/genética , Regulação Neoplásica da Expressão Gênica , Fator de Crescimento de Hepatócito/genética , Proteínas de Homeodomínio/genética , Neoplasias Mamárias Experimentais/tratamento farmacológico , Neovascularização Patológica/prevenção & controle , Proteínas Nucleares/genética , Ácidos Fosfatídicos/farmacologia , Proteínas Repressoras/genética , Animais , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Embrião de Galinha , Membrana Corioalantoide/irrigação sanguínea , Membrana Corioalantoide/efeitos dos fármacos , Feminino , Fator 1 de Crescimento de Fibroblastos/metabolismo , Fator de Crescimento de Hepatócito/metabolismo , Proteínas de Homeodomínio/metabolismo , Células Endoteliais da Veia Umbilical Humana/citologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Neoplasias Mamárias Experimentais/genética , Neoplasias Mamárias Experimentais/metabolismo , Neoplasias Mamárias Experimentais/patologia , Camundongos , Camundongos Nus , Neovascularização Patológica/genética , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Proteínas Nucleares/metabolismo , Proteínas Repressoras/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fatores de Transcrição , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Neoplasias de Mama Triplo Negativas/genética , Neoplasias de Mama Triplo Negativas/metabolismo , Neoplasias de Mama Triplo Negativas/patologia , Carga Tumoral/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
12.
Arch Biochem Biophys ; 654: 115-125, 2018 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-30031837

RESUMO

Human acidic fibroblast growth factor 1 (hFGF1) is a protein intricately involved in cell growth and tissue repair. In this study, we investigate the effect(s) of understanding the role of a conserved proline (P135), located in the heparin binding pocket, on the structure, stability, heparin binding affinity, and cell proliferation activity of hFGF1. Substitution of proline-135 with a positively charged lysine (P135K) resulted in partial destabilization of the protein; however, the overall structural integrity of the protein was maintained upon substitution of proline-135 with either a negative charge (P135E) or a polar amino acid (P135Q). Interestingly, upon heparin binding, an increase in thermal stability equivalent to that of wt-hFGF1 was observed when P135 was replaced with a positive (P135K) or a negative charge (P135E), or with a polar amino acid (P135Q). Surprisingly, introduction of negative charge in the heparin-binding pocket at position 135 (P135E) increased hFGF1's affinity for heparin by 3-fold, while the P135K mutation, did not alter the heparin-binding affinity. However, the enhanced heparin-binding affinity of mutant P135E did not translate to an increase in cell proliferation activity. Interestingly, the P135K and P135E double mutations, P135K/R136E and P135/R136E, reduced the heparin binding affinity by ∼3-fold. Furthermore, the cell proliferation activity was increased when the charge reversal mutation R136E was paired with both P135E (P135E/R136E) and P135K (P135K/R136E). Overall, the results of this study suggest that while heparin is useful for stabilizing hFGF1 on the cell surface, this interaction is not mandatory for activation of the FGF receptor.


Assuntos
Proliferação de Células/fisiologia , Fator 1 de Crescimento de Fibroblastos/química , Fator 1 de Crescimento de Fibroblastos/fisiologia , Prolina/fisiologia , Fator 1 de Crescimento de Fibroblastos/genética , Heparina/metabolismo , Humanos , Mutagênese Sítio-Dirigida , Ligação Proteica , Estabilidade Proteica , Estrutura Terciária de Proteína , Espectroscopia de Prótons por Ressonância Magnética , Receptores de Fatores de Crescimento de Fibroblastos/metabolismo
13.
Mol Med Rep ; 18(1): 1025-1030, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29845277

RESUMO

Atherosclerosis is recognized at present as a chronic metabolic disease of the arteries that leads to multifocal plaque development. Previous studies have reported that acid fibroblast growth factor (aFGF) is a critical therapeutic regulator in numerous chronic metabolic disorders. However, there is currently no direct evidence indicating whether aFGF serves a therapeutic role in atherosclerosis. In the present study, the role of aFGF in atherosclerotic lesion development was investigated by examining high­fat diet (HFD)­fed apolipoprotein E (ApoE)­/­ mice and parenteral administration of aFGF. Increased expression of aFGF and peroxisome proliferator­activated receptor α (PPARα) was observed during atherosclerotic lesion development. The parenteral delivery of aFGF facilitated the progression of atherosclerosis without altering serum lipid expression levels in HFD­fed ApoE­/­ mice. Furthermore, it was demonstrated that aFGF increased the expression of PPARα and inflammatory cytokines. The present results provided evidence that aFGF accelerates the progression of atherosclerosis and suggested that aFGF may be a potential therapeutic target for the prevention of atherosclerosis development.


Assuntos
Apolipoproteínas E/deficiência , Aterosclerose , Gorduras na Dieta/efeitos adversos , Fator 1 de Crescimento de Fibroblastos , Placa Aterosclerótica , Animais , Aterosclerose/induzido quimicamente , Aterosclerose/genética , Aterosclerose/metabolismo , Aterosclerose/patologia , Gorduras na Dieta/farmacologia , Fator 1 de Crescimento de Fibroblastos/genética , Fator 1 de Crescimento de Fibroblastos/metabolismo , Camundongos , Camundongos Knockout , PPAR alfa/genética , PPAR alfa/metabolismo , Placa Aterosclerótica/induzido quimicamente , Placa Aterosclerótica/genética , Placa Aterosclerótica/metabolismo , Placa Aterosclerótica/patologia
14.
Proc Natl Acad Sci U S A ; 115(20): 5235-5240, 2018 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-29695630

RESUMO

Reef-building corals are critically important species that are threatened by anthropogenic stresses including climate change. In attempts to understand corals' responses to stress and other aspects of their biology, numerous genomic and transcriptomic studies have been performed, generating a variety of hypotheses about the roles of particular genes and molecular pathways. However, it has not generally been possible to test these hypotheses rigorously because of the lack of genetic tools for corals. Here, we demonstrate efficient genome editing using the CRISPR/Cas9 system in the coral Acropora millepora We targeted the genes encoding fibroblast growth factor 1a (FGF1a), green fluorescent protein (GFP), and red fluorescent protein (RFP). After microinjecting CRISPR/Cas9 ribonucleoprotein complexes into fertilized eggs, we detected induced mutations in the targeted genes using changes in restriction-fragment length, Sanger sequencing, and high-throughput Illumina sequencing. We observed mutations in ∼50% of individuals screened, and the proportions of wild-type and various mutant gene copies in these individuals indicated that mutation induction continued for at least several cell cycles after injection. Although multiple paralogous genes encoding green fluorescent proteins are present in A. millepora, appropriate design of the guide RNA allowed us to induce mutations simultaneously in more than one paralog. Because A. millepora larvae can be induced to settle and begin colony formation in the laboratory, CRISPR/Cas9-based gene editing should allow rigorous tests of gene function in both larval and adult corals.


Assuntos
Sistemas CRISPR-Cas , Recifes de Corais , Fator 1 de Crescimento de Fibroblastos/antagonistas & inibidores , Edição de Genes , Proteínas de Fluorescência Verde/antagonistas & inibidores , Proteínas Luminescentes/antagonistas & inibidores , Mutação , Animais , Sequência de Bases , Fator 1 de Crescimento de Fibroblastos/genética , Genoma , Genômica , Proteínas de Fluorescência Verde/genética , Proteínas Luminescentes/genética , Fenótipo , Homologia de Sequência
15.
Clin Cancer Res ; 24(16): 3888-3897, 2018 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-29691299

RESUMO

Purpose: A comprehensive analysis of the genomics of undifferentiated sarcomas (UDS) is lacking. We analyzed copy-number alterations and fusion status in patients with UDS prospectively treated on Children's Oncology Group protocol ARST0332.Experimental Design: Copy-number alterations were assessed by OncoScan FFPE Express on 32 UDS. Whole-exome and transcriptome libraries from eight tumors with sufficient archived material were sequenced on HiSeq (2 × 100 bp). Targeted RNA-sequencing using Archer chemistry was performed on two additional cases.Results: Five-year overall survival for patients with UDS was 83% (95% CI, 69%-97%) with risk-adapted therapy (surgery, chemotherapy, and radiotherapy). Both focal and arm-level copy-number alterations were common including gain of 1q (8/32, 25%) and loss of 1p (7/32, 22%), both of which occurred more often in clinically defined high-risk tumors. Tumors with both loss of 1p and gain of 1q carried an especially poor prognosis with a 5-year event-free survival of 20%. GISTIC analysis identified recurrent amplification of FGF1 on 5q31.3 (q = 0.03) and loss of CDKN2A and CDKN2B on 9p21.3 (q = 0.07). Known oncogenic fusions were identified in eight of 10 cases analyzed by next-generation sequencing.Conclusions: Pediatric UDS generally has a good outcome with risk-adapted therapy. A high-risk subset of patients whose tumors have copy-number loss of 1p and gain of 1q was identified with only 20% survival. Oncogenic fusions are common in UDS, and next-generation sequencing should be considered for children with UDS to refine the diagnosis and identify potentially targetable drivers. Clin Cancer Res; 24(16); 3888-97. ©2018 AACR.


Assuntos
Tratamento Farmacológico , Radioterapia , Sarcoma/genética , Sarcoma/terapia , Adolescente , Adulto , Biomarcadores Tumorais/genética , Diferenciação Celular/genética , Criança , Pré-Escolar , Aberrações Cromossômicas , Terapia Combinada , Inibidor de Quinase Dependente de Ciclina p15/genética , Inibidor p16 de Quinase Dependente de Ciclina/genética , Variações do Número de Cópias de DNA/genética , Feminino , Fator 1 de Crescimento de Fibroblastos/genética , Dosagem de Genes , Predisposição Genética para Doença , Humanos , Lactente , Estimativa de Kaplan-Meier , Masculino , Proteínas de Fusão Oncogênica/genética , Intervalo Livre de Progressão , Sarcoma/classificação , Sarcoma/patologia , Transcriptoma/genética , Sequenciamento Completo do Exoma , Adulto Jovem
16.
J Biomed Sci ; 25(1): 18, 2018 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-29490650

RESUMO

BACKGROUND: LncRNA Gas5 is known to be a key control element during growth, differentiation and development in mammalian species. However, the role and function of Gas5 in growth plate chondrocytes has not been determined. METHODS: The overexpression and knockdown models of Gas5 and miR-21 in cells and animals were constructed. Cell survival was determined by MTT assay and flow cytometry. Animal biochemical indices were measured by enzyme-linked immunosorbent assay, hematoxylin/eosin staining, immunohistochemistry or in situ hybridisation. Dual luciferase reporter gene assay was carried out to study targeting. RESULTS: First, we found the expression levels of fibroblast growth factor 1(FGF1) were up-regulated and miR-21 were down-regulated in Gas5 overexpressing model cells. Meanwhile, the expression levels of FGF1 and Gas5 were up-regulated in miR-21 knockdown model cells. Furthermore, cell proliferation was significantly promoted after Gas5 knockdown or miR-21 overexpression. Subsequently, Gas5 promoted apoptosis, while miR-21 suppressed apoptosis. Animal assays demonstrated that both Gas5 and dexamethasone suppressed proliferation and promoted apoptosis of growth plate chondrocytes, up-regulated FGF1 expression but reduced miR-21 expression. Finally, there was a binding relationship between Gas5, miR-21 and FGF1. CONCLUSION: We concluded that Gas5 regulated proliferation and apoptosis in growth plate by controlling FGF1 expression via miR-21 regulation.


Assuntos
Fator 1 de Crescimento de Fibroblastos/genética , Regulação Neoplásica da Expressão Gênica/genética , MicroRNAs/genética , RNA Longo não Codificante/genética , Animais , Apoptose/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Condrócitos/fisiologia , Fator 1 de Crescimento de Fibroblastos/metabolismo , Lâmina de Crescimento/fisiologia , Humanos , Masculino , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , Ratos , Ratos Sprague-Dawley , Organismos Livres de Patógenos Específicos
17.
J Cell Mol Med ; 22(6): 3259-3263, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29575613

RESUMO

Single-chain variable fragment (scFv) antibodies are the smallest immunoglobulins with high antigen-binding affinity. We have previously reported that fibroblast growth factor 1 played pivotal roles in cancer development and generated a mouse scFv (mscFv1C9) could effectively prohibit cancer cell proliferation in vitro and in vivo. Here, we further humanized this scFv (hscFv1C9) using a structure-guided complementarity determining region grafting strategy. The purified hscFv1C9 maintained similar antigen-binding affinity and specificity as mscFv1C9, and it was capable of inhibiting growth of different tumours in vitro and in vivo. These data strongly suggested that hscFv1C9 has antitumour potentials.


Assuntos
Neoplasias da Mama/tratamento farmacológico , Fator 1 de Crescimento de Fibroblastos/antagonistas & inibidores , Glioma/tratamento farmacológico , Anticorpos de Cadeia Única/farmacologia , Sequência de Aminoácidos/genética , Animais , Anticorpos Monoclonais Humanizados/imunologia , Anticorpos Monoclonais Humanizados/farmacologia , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Feminino , Fator 1 de Crescimento de Fibroblastos/química , Fator 1 de Crescimento de Fibroblastos/genética , Fator 1 de Crescimento de Fibroblastos/imunologia , Glioma/genética , Glioma/patologia , Xenoenxertos , Humanos , Camundongos , Anticorpos de Cadeia Única/imunologia
18.
J Comp Neurol ; 526(7): 1195-1208, 2018 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-29405296

RESUMO

Nerves are particularly vulnerable to damage due to their unique structure with meter-long axons. In the peripheral nervous system neurons and Schwann cells can activate the injury-response program that directs axons to either regenerate or degenerate after traumatic nerve injury. However, the differences between the genetic programs driving nerve regeneration and degeneration have not yet been described extensively. To understand these differences, in this study we have compared the injury-induced transcriptomic changes between the regenerating proximal segment and the degenerating distal segment of a transected nerve, at different post-injury time points. We analyzed the spatiotemporal dynamics of the mouse transcriptome using a sciatic nerve-injury model by means of RNA sequencing. The results of the differentially regulated genes (DEGs) analysis show that some DEG groups are similarly regulated in both proximal and distal segments, and primarily display a positive correlation. However, some DEG groups are exclusively regulated in either the proximal or the distal segment, suggesting that these DEG groups constitute a genetic network for distinguishing the regenerative and degenerative responses. In addition, our gene ontology analysis revealed an enrichment of particular biological processes in different phases and locations. Thus, our data provide a spatiotemporal profile of the transcriptomes that are differentially regulated in either regenerating or degenerating nerves, in vivo. The specific biological processes enriched in the DEG groups might delineate the injury-responsive program that induces contrasting regenerative and degenerative responses in different nerve segments.


Assuntos
Regulação da Expressão Gênica/fisiologia , Neuropatia Ciática/patologia , Neuropatia Ciática/fisiopatologia , Transcriptoma/fisiologia , Animais , Quimiocina CCL2/metabolismo , Modelos Animais de Doenças , Feminino , Fator 1 de Crescimento de Fibroblastos/genética , Fator 1 de Crescimento de Fibroblastos/metabolismo , Redes Reguladoras de Genes , Cinesina/genética , Cinesina/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , RNA Mensageiro/metabolismo , Fatores de Tempo
19.
Brain Res Bull ; 141: 3-12, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29477835

RESUMO

Recent evidence demonstrates that epigenetic regulation of gene transcription is critically involved in learning and memory. Here, we discuss the role of histone acetylation and DNA methylation, which are two best understood epigenetic processes in memory processes. More specifically, we focus on learning-strength-dependent changes in chromatin on the fibroblast growth factor 1 (Fgf1) gene and on the molecular events that modulate regulation of Fgf1 transcription, required for memory enhancement, with the specific focus on CREB-regulated transcription coactivator 1 (CRTC1).


Assuntos
Epigênese Genética , Fator 1 de Crescimento de Fibroblastos/metabolismo , Memória/fisiologia , Fatores de Transcrição/metabolismo , Animais , Encéfalo/metabolismo , Fator 1 de Crescimento de Fibroblastos/genética , Regulação da Expressão Gênica/fisiologia , Humanos
20.
Mol Neurobiol ; 55(4): 3408-3425, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28502041

RESUMO

Glucose is believed to improve the memory in both human and mice, but the detailed insights were mostly elusive. In this study, we focused on two major neurotrophic factors, brain-derived neurotrophic factor (BDNF) and fibroblast growth factor 1 (FGF1), which are believed to be associated with the memory enhancement and assessed their expressional regulation among the murine neuronal and glial cells. Our findings showed that the glucose administration increased phosphorylated Akt, phosphorylated CREB, exon 1- and exon 4-specific BDNF transcripts, and FGF1 transcripts that are associated with the epigenetic changes expected to open the chromatin and a reduction in histone deacetylase 2 (HDAC2) in neurons and astrocytes of the murine hippocampus. The glucose administration enhanced the long-term potentiation and the number of dendritic spines in the CA1 and CA3 subfields of hippocampus. The intrahippocampal injection of short hairpin RNA against TrkB canceled the glucose-mediated memory enhancement. Like the glucose, we also report that the HDAC inhibitor can enhance the memory through the BDNF-TrkB pathway but it targeted different brain cell populations to enhance the BDNF and FGF1 transcripts. In addition, the soluble FGF1 treatments significantly increased the BDNF expression in astrocytes and neurons, suggesting that the glucose-mediated induction of the neurotrophic factors could contribute to the memory. Our study provides the valuable insights, explaining the distinctive neuronal and glial cell regulation of the neurotrophic factors by glucose and HDAC inhibitor, which could likely explain how our brain cells can control the release of neurotrophic factors.


Assuntos
Fator Neurotrófico Derivado do Encéfalo/genética , Encéfalo/citologia , Epigênese Genética/efeitos dos fármacos , Fator 1 de Crescimento de Fibroblastos/genética , Glucose/farmacologia , Acetilação , Animais , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Espinhas Dendríticas/efeitos dos fármacos , Espinhas Dendríticas/metabolismo , Retroalimentação Fisiológica , Fator 1 de Crescimento de Fibroblastos/metabolismo , Hipocampo/citologia , Inibidores de Histona Desacetilases/farmacologia , Histonas/metabolismo , Potenciação de Longa Duração/efeitos dos fármacos , Memória/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Fosforilação/efeitos dos fármacos , Regiões Promotoras Genéticas/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor trkB/metabolismo , Vorinostat/farmacologia
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