Análisis de microdensidad vascular y factores de crecimiento en carcinoma oral de células escamosas / Analysis of vascular microdensity and growth factors in oral squamous cell carcinoma

Introducción: El carcinoma oral de células escamosas (COCE) es una neoplasia epitelial maligna que se presenta frecuentemente entre la quinta y sexta década de la vida. Su compleja patogénesis incluye el proceso de angiogénesis y la regulación del microambiente tumoral como mecanismos de progresión tumoral. Objetivo: Determinar la relación entre las variables clínicas e histológicas del COCE con la inmunoexpresión de VEGF, FGF-1, FGFR-1, TGFB-1, TGFBR-II y CD105. Material y métodos: Nueve casos de COCE; tres bien (BD), tres moderado (MD) y tres pobremente diferenciados (PD) obtenidos del Departamento de Patología y Medicina Bucal, División de Estudios de Postgrado e Investigación. Se aplicó la técnica de inmunohistoquímica por peroxidasa para identificar la expresión de VEGF, FGF-1, FGFR- 1, TGFB-1, TGFBR-II y CD105. El análisis de inmunoexpresión se realizó con el programa ImageJ. Se aplicó la prueba de Kruskal-Wallis y correlación de Spearman (p < 0.05). Resultados: La inmunoexpresión de VEGF fue mayor en los COCE PD, FGFR-1 fue positivo en los BD, mientras que FGF, TGFB-1 y TGFBR-II fueron negativos. El análisis de microdensidad vascular (MVD) indicó mayor número de vasos CD105 positivos en los carcinomas BD, seguidos de los PD y MD. Conclusión: Considerando los resultados obtenidos podemos concluir que la angiogénesis es un fenómeno constante independiente del grado de diferenciación que durante el proceso de transformación de una neoplasia requerirá la formación de vasos sanguíneos y que este proceso puede ser modulado por factores de crecimiento tales como los analizados en este trabajo (AU)

Introduction: Oral squamous cell carcinoma (OSCC) is a malignant epithelial neoplasm that frequently occurs between the fifth and sixth decade of life. Its complex pathogenesis includes the angiogenesis process and the regulation of the tumor microenvironment as mechanisms of tumor progression. Objective: To determine the relationship between the clinical and histological variables of OSCC with the immunoexpression of VEGF, FGF-1, FGFR-1, TGFB- 1, TGFBR-II and CD105. Material and methods: Nine cases of OSCC; three well (WD), three moderate (MD) and three poorly differentiated (PD) obtained from the Oral Medicine and Pathology Department, Division of Graduate Studies and Research. The peroxidase immunohistochemistry technique was performed to identify the expression of VEGF, FGF-1, FGFR-1, TGFB-1, TGFBR-II and CD105. The immunoexpression analysis was performed with the ImageJ software. The Kruskal-Wallis and Spearman correlation test were performed (p < 0.05). Results: VEGF immunoexpression was higher in PD OSCC, while FGFR-1 was predominantly positive in WD; FGF, TGFB-1 and TGFBR-II were negative. Vascular microdensity analysis (MVD) indicated a greater number of CD105 positive vessels in WD carcinomas, followed by PD and MD. Conclusion: Considering the results obtained, we can conclude that angiogenesis is a constant phenomenon independent of the degree of differentiation; that during the transformation process of a neoplasm it will require the formation of blood vessels and that this process can be modulated by growth factors such as those analyzed in this work (AU)

FGFR1 is an independent prognostic factor and can be regulated by miR-497 in gastric cancer progression

Fibroblast growth factor receptor 1 (FGFR1) has been reported in gastric cancer to be a prognostic factor. However, miR-497-targeted FGFR1 has not been explored in the carcinogenesis of gastric cancer. The present study intended to revalidate the prognostic significance of FGFR1 in patients with gastric cancer, and the mechanism of miR-497-regulated FGFR1 was investigated in gastric cancer cell proliferation and apoptosis. The messenger RNA (mRNA) and protein levels were assayed by RT-qPCR and western blotting, respectively. The targeted genes were predicted by a bioinformatics algorithm and confirmed by a dual luciferase reporter assay. Cell proliferation was analyzed by CCK-8 assay. Annexin V-FITC/PI staining was used to evaluate the apoptosis in AGS and SGC-7901 cells. FGFR1 was frequently up-regulated in gastric cancer tissues and associated with poor overall survival in patients with gastric cancer. Interestingly, FGFR1 loss-of-function resulted in a significant growth inhibition and apoptosis in AGS and SGC-7901 cells. In addition, we found that miR-497 was inhibited in gastric cancer tissues and cell lines, while overexpression of miR-497 could suppress proliferation and induce apoptosis in AGS and SGC-7901 cells. Importantly, bioinformatics analysis and experimental data suggested that FGFR1 was a direct target of miR-497, which could inhibit FGFR1 expression when transfected with miR-497 mimics. Furthermore, we found that overexpression of FGFR1 reversed the growth inhibition and apoptosis of miR-497 mimics in AGS and SGC-7901 cells. These findings suggested that overexpression of miR-497 inhibited proliferation and induced apoptosis in gastric cancer through the suppression of FGFR1.

Inhibitory Eefects of the novel tyrosine kinase inhibitor BGJ398 against human leukemic cell line KG-1 cells / 中华血液学杂志

Objective@#To explore the effects and possible mechanisms of the novel pan-FGFR inhibitor BGJ398 on KG-1 cells in vitro.@*Methods@#Effects of BGJ398 on cells proliferation were detected by CCK-8, the apoptosis was assessed by Annexin V-FITC. Reverse transcriptionquantitative polymerase chain reaction (q-PCR) analysis was used to detect the expression of apoptosis-related genes B cell lymphoma-2 (Bcl-2) and caspase-3. Western blotting analysis was performed to explore the proteins expression levels of Bcl-2, caspase-3 and the expression of p-AKT, p-S6K, p-ERK and FGFR1.@*Results@#BGJ398 effectively inhibited cell proliferation by dose-dependent manners. BGJ398(1.4 µmol/L) induced apoptosis of KG-1 cells by 36.4%, compared with 4.5% in the control group(P<0.001). Treatment with BGJ398 at 1.4 µmol/L led to significant increases in the expression levels of caspase-3, and decreases in the expression of Bcl-2 (P<0.005). In accordance with these results, Western blot analysis further confirmed the increased expression of Bcl-2 protein along with elevated caspase-3 activity. In addition, BGJ398 markedly down-regulated FGFR1OP2-FGFR1 fusion protein, p-AKT and p-S6K expression, but not p-ERK expression.@*Conclusion@#Novel pan-FGFR inhibitor BGJ398 substantially suppressed KG-1 cell growth and induced apoptosis by inhibiting the expression of FGFR1, p-AKT, p-S6K and regulating apoptosis-related proteins.

Fibroblast growth factor receptor-1 is essential for regulation of mitochondrial biogenesis via control of microRNA let-7b / 中华内分泌代谢杂志

Objective To explore the role of fibroblast growth factor receptor ( FGFR ) 1 in endothelial homeostasis via an induction of microRNA let-7s, with effects on AcSDKP(N-acetyl-seryl-aspartyl-lysyl-proline) and associated mitochondrial biogenesis. Methods Blocking FGFR1 signaling pathway, Western blot and immunofluorescence staining were used to measure mitochondrial fusion ( mitofusin-2, MFN2;optic atrophy protein 1, OPA1 ) and fission ( dynamin-related protein-1, DRP1 ) proteins and mitochondrial biogenesis by MitoTraker Green. Also real-time quantitative PCR(qPCR) was performed to test microRNA let-7' expression. Results FGFR1 signaling pathway was critical for AcSDKP maintaining mitochondrial biogenesis through induction of microRNA let-7b. In endothelial cells, the AcSDKP restored the triple[TGF-β2, interleukin (IL)-1β, tumor necrosis factor (TNF)-α]-suppressed microRNA let-7b-5p expression and associated with mitochondrial biogenesis. Such effect of AcSDKP was lost in either fibroblast growth factor receptor substrate 2 (FRS2) siRNA or neutralizing FGFR1 treated-cells. Similarly, AcSDKP lost its effect on mitochondrial biogenesis in microRNA let-7b-5p inhibitor-treated-cells. In addition, microRNA let-7b-5p mimic reversed the FRS2 siRNA-suppressed mitochondrial biogenesis in endothelial cells. Conclusion These findings demonstrated that FGFR1 is critical for maintaining mitochondrial biogenesis through control of microRNA let-7b-5p in endothelial cells.

Association Study of Fibroblast Growth Factor 2 and Fibroblast Growth Factor Receptors Gene Polymorphism in Korean Ossification of the Posterior Longitudinal Ligament Patients

OBJECTIVE: The aim of this study was to determine whether single nucleotide polymorphisms (SNPs) of fibroblast growth factor (FGF) 2 gene and fibroblast growth factor receptor (FGFR) genes are associated with ossification of the posterior longitudinal ligament (OPLL). METHODS: A total of 157 patients with OPLL and 222 controls were recruited for a case control association study investigating the relationship between SNPs of FGF2, FGFR1, FGFR2 and OPLL. To identify the association among polymorphisms of FGF2 gene, FGFR1, FGFR2 genes and OPLL, the authors genotyped 9 SNPs of the genes (FGF2 : rs1476217, rs308395, rs308397, and rs3747676; FGFR1 : rs13317 and rs2467531; FGFR2 : rs755793, rs1047100, and rs3135831) using direct sequencing method. SNPs data were analyzed using the SNPStats, SNPAnalyzer, Haploview, and Helixtree programs. RESULTS: Of the SNPs, a SNP (rs13317) in FGFR1 was significantly associated with the susceptibility of OPLL in the codominant (odds ratio=1.35, 95% confidence interval=1.01-1.81, p=0.048) and recessive model (odds ratio=2.00, 95% confidence interval=1.11-3.59, p=0.020). The analysis adjusted for associated condition showed that the SNP of rs1476217 (p=0.03), rs3747676 (p=0.01) polymorphisms in the FGF2 were associated with diffuse idiopathic skeletal hyperostosis (DISH) and rs1476217 (p=0.01) in the FGF2 was associated with ossification of the ligament flavum (OLF). CONCLUSION: The results of the present study revealed that an FGFR1 SNP was significantly associated with OPLL and that a SNP in FGF2 was associated with conditions that were comorbid with OPLL (DISH and OLF).

Clinical Significance of FGFR1OP and p57 (Kip2) Proteins in Gliomas: An Immunohistochemical Study / 中国肿瘤临床

Objective: To investigate the effects of FGFR1OP and p57/Kip2 proteins on the genesis and progression of gliomas and their clinical significance. Methods: The expression of FGFR1OP and p57/Kip2 in 54 glioma specimens was detected by SP immunohistochemical technique. The relationship between the ex-pression levels of those proteins and various clinical pathologic factors was evaluated. Results: The expres-sion of FGFR1OP and p57/Kip2 was found in 66.7% and 44.4% gliomas, respectively. The OD value of FG-FR1OP was 0.131±0.010 in high grade gliomas, and 0.118±0.010 in low grade ones, with a statistical signifi-cance (t=-5.497, P=0.000), showing that higher expression of FGFR1OP was significantly associated with glo-ma cell differentiation. The OD value of p57/Kip2 was 0.156±0.008 in high grade gliomas, and 0.165±0.006 in low grade ones, with a statistical significance (t=0.296, P=0.014), showing that lower p57/Kip2 expression was correlated with high grade gliomas. FGFR1OP was negatively correlated with p57/Kip2 in gliomas (r=-0.732, P<0.01). Conclusion: Increased expression of FGFR1OP and/or decreased expression of p57/Kip2 may play an important role in the genesis and progression of gliomas and may indicate a poor prognosis.

Síndrome de Kallmann: uma revisão histórica, clínica e molecular / Kallmann syndrome: a hystorical, clinical and molecular review

A síndrome de Kallmann (SK) é a associação de hipogonadismo hipogonadotrófico (HH) e anosmia descrita por Maestre de San Juan, em 1856, e caracterizada como condição hereditária por Franz Josef Kallmann, em 1944. Muitos aspectos de sua patogenia, variabilidade fenotípica e genotípica foram desvendados nos últimos 15 anos. Conseqüentemente, tem sido difícil manter-se atualizado frente à rapidez que o conhecimento dessa condição é gerado. Nesta revisão, resgatamos aspectos históricos pouco conhecidos sobre a síndrome e seus descobridores; incorporamos novas descobertas relacionadas à embriogênese dos neurônios olfatórios e produtores de GnRH. Esse processo é fundamental para compreender a associação de hipogonadismo e anosmia; descrevemos a heterogeneidade fenotípica e genotípica, incluindo mutações em cinco genes (KAL-1, FGFR1, PROKR2, PROK2 e NELF). Para cada gene, discutimos a função da proteína codificada na migração e maturação dos neurônios olfatórios e GnRH a partir de estudos in vitro e modelos experimentais e descrevemos características clínicas dos portadores dessas mutações.

Kallmann syndrome (KS), the association of hypogonadotropic hypogonadism and anosmia, was described by Maestre de San Juan in 1856 and characterized as a hereditary condition by Franz Josef Kallmann in 1944. Many aspects such as pathogeny, phenotype and genotype in KS were described in the last fifteen years. The knowledge of this condition has grown fast, making it difficult to update. Here we review historical aspects of this condition and its discoverers and describe new findings regarding the embryogenesis of the olfactory bulb and GnRH secreting neuronal tracts that are important for understanding the association of hypogonadism and anosmia. Additionally, we describe the phenotypic and genotypic heterogeneity of KS, including five related genes (KAL-1, FGFR1, PROKR2, PROK2 e NELF), and discuss the function of each codified protein in migration and maturation of the olfactory and GnRH neurons, with data from in vitro and in vivo studies. Finally we describe the clinical phenotype of patients carrying these mutations.

Molecular Genetic Analysis and Endocriological Evaluation of Isolated Hypogonadotropic Hypogonadism Presented with Delayed Puberty / 대한소아내분비학회지

PURPOSE: Isolated hypogonadotropic hypogonadism (HH) is a disorder of the hypothalamic-pituitary axis causing gonadotrophin releasing hormone or gonadotrophin deficiency. Kallman syndrome has a constellation of features, characterized by HH, hyposomia, deafness and congenital heart defects, whereas a normal sense of smell indicates idiopathic HH. Defects in some genes such as KAL-1, FGFR1, GNRHR and GPR54, have been described. The study was undertaken to identify the molecular defects of aforementioned genes and to evaluate clinical profiles in patients with HH. METHODS: Among the patients who visited the hospital due to delayed puberty from March 1995 to March 2006, seven male patients were suspected of having HH and included in this study. Clinical characteristics at diagnosis and endocrinological data including gonadotropin-releasing hormone (GnRH) agonist stimulation test were investigated. For molecular genetic evaluation, routine karyotyping in all patients and mutation analyses of the KAL, FGFR-1, GNRHR and GPR54 genes in six patients were performed. RESULTS: The study included 7 patients diagnosed as HH by GnRH stimulation; 4 with Kallman syndrome and 3 with idiopathic HH. No mutation was identified by DNA sequence analysis of KAL-1, FGFR1, GNRHR and GPR54 genes in 7 patients with HH. At diagnosis, chronologic age was 16.88+/-.90 years; height SDS, -0.36+/-.43; mean volume of the testis, 1.79+/-.76 mL. Of the patients with Kallman syndrome, 3 had sensory neural hearing loss, 2 congenital heart disease and 1 bilateral cryptorchidism. The olfactory bulb or sulci hypoplasia was found in all Kallman syndrome patients on the brain MRI. No abnormal finding was found in the brain MRI of the patients with idiopathic HH. Peak LH and FSH levels were 1.27+/-.60 IU/L and 1.15+/-.65 IU/L after GnRH stimulation. Baseline total testosterone level was 0.41+/-.24 ng/mL. The patients were treated with testosterone enanthate every 3-4 weeks for the mean duration of 40.60+/-8.61 months. During the follow-up period, 5 patients reached the final adult height with the mean height of 175.00+/-.47 cm (0.28+/-.14 SDS). CONCLUSION: For differential diagnosis of delayed puberty, physical, radiological, hormonal evaluations are all necessary. Many genes associated with Isolated HH were founded until now. But, mutations in these genes account for small proportion of Isolated HH yet. Further study of genes that regulate secondary sexual development and function will give important information regarding the development of normal puberty in humans.

Expression and significance of basic fibroblast gwowth factor and fibroblast growth factor receptor-1 in ovarian epithelial neoplasm / 西安交通大学学报·英文版

Objective: To study the relevance of expression of basic fibroblast growth factor (bFGF), fibroblast growth factor receptor-1 (FGFR-1) and carcinogenesis and progression of ovarian epithelial neoplasm. Methods: Ten cases of normal ovarian tissues and 75 cases of ovarian epithelial neoplasm tissues were detected by immunohistochemical methods: S-P for bFGF, FGFR-1, double immunohistochemistry Lab-SA for Ki-67 antigen and bFGF. Results: The expression level of bFGF, FGFR-1 in ovarian epithelium and ovarian epithelial neoplasm showed a step-wise increase in the following order: normal 〈benign 〈borderline 〈malignant; The expression level and intensity of bFGF and FGFR-1 were increased with the decrease of differentiation degree and increase of clinical stage in ovarian carcinoma; There was no statistical difference between the expression of bFGF, FGFR-1 in serous cystadenocarcinoma and that of mucinous cystadenocarcinoma; The expression of bFGF was correlated with that of FGFR-1 in neoplastic tissues; There were positive expression rates of bFGF and Ki-67 antigen in ovarian epithelial neoplasm. Conclusion: As an important proliferative factor, bFGF plays an important role in carcinogenisis and progression of ovarian epithelial neoplasm.

Changes of type Ⅰ fibroblast growth factor receptor gene during development / 免疫学杂志

Objective To find the changes of human fibroblast growth factor receptor 1 genone during development.Method Southern blot analysis of genomic DNA isolated from adult and fetal tissues. Result Adult FGFR1 gene structure is different from its embryonic counterpart. Conclusion The disserence might lead to changes of FGFR1 expression as wel as functions of the cells.

Level of bFGF and its receptor FGFR1 mRNA in rat brain following injury caused by moderate lateral fluid percussion / 中国法医学杂志

Objective Study on the pattern of changes of bFGF and FGFRl mRNA occurred in the experimental brain injury model in order to provide scientific basis for the diagnosis, forensic identification and clinical treatment, and also for further ascertaining the molecular mechanism of brain injury. Methods Male Sprague-Dawley rats were divided into 3 groups: normal control, sham operation, and injury groups. The rats of injury groups were subjected to moderate lateral fluid percussion brain injury (0.2MPa). The injury groups was then subdivided into 30min, 1h, 3h, 6h, 12h, 1d, 3d and 7d groups according to the time elapsed after injury. In situ hybridization (ISH) and RT-PCR were used for studying the mRNA expression of both bFGF and FGFRl factors. Results (1) In the brain of normal control and sham operation control groups, mRNA levels of bFGF and FGFRl were low; (2) There is gradual increase of bFGF and FGFRl mRNA levels could be observed 6h to 3d after injury both in cortex and brain stem, then partly declined at 7d; (3) In hippocampus, the gradual increase occurred during 3h- 1d after injury, then partly declined at 3d, and returned to basal level at 7d. Conclusions The results suggested that brain injury induced the gene expressions of bFGF and FGFR1. The bFGF may contribute to maintenance of nerve cell survival and the repair of damaged neural tissues after CNS injury and the patterns of their level change were quite regular. It is potentially useful for timing of injury in forensic medical practice.

The expression of human fibroblast growth factor receptor 1 on the membrane of insect cell Sf9 / 中国免疫学杂志

Objective:Express the recombinant human FGFR1 in order to screen the FGFs.Methods:A cDNA fragment encoding human fibroblast growth factor receptor 1(FGFR1) was isolated by RT PCR from human lung fibroblasts,and then cloned into pCR TM II plasmid and pFastBac1 donor plasmid.Through transposition,recombinant bacmid FGFR1 DNA was formed and was then transfected into Sf9 insect cells to produce recombinant Baculovirus.Sf9 insect cell were infected with recombinant Baculovirus.The harvested culture supernatant was subjected to Western Blot and ELISA analysis.Results:The size of FGFR1 cDNA fragment is 2 100 bp.The MW of expressed recombinant FGFR1 was 78 kD.ELISA showed that human recombinant FGFR1 was expressed on the membrane of insect cell Sf9.Conclusion:The recombinant human FGFR1 can be expressed on the membrane of insect cell Sf9 effectively.

Effects of antisense oligonucleotide on endogenous human bFGF, bFGF mRNA and FGFR1 in tumor cell SWO-38 / 中国免疫学杂志

Objective:To clarify the contribution of endogenous bFGF, bFGF mRNA and FGFR1 to the abnormal growth and phenotypic transformation of neoplastic tumors cells.Methods:The antisense oligonucleotide primers was used to evaluate the influence of endogenous bFGF on growth of human glioma malignant cell lines SWO-38 in vitro. MTT was used to examine the variety of cells growth treated with bFGF antisense oligonucleotide primers. The methods of ELISA, in situ hybridization, immuno-hischemistry and image analysis were used to detect the expression level of bFGF, bFGF mRNA and FGFR1. The colony formation of cells in soft agar was used to assess the cloning efficiency of the cells after exposed to bFGF antisense oligo-nucleotide primers.Results:The cells multiplication, expression of bFGF mRNA and FGFR1 was inhibited by bFGF antisense oligonucleotide primers,and the cells multiplication was dose-dependent. Treated with antisense oligo-nucleotide primers, the expression of FGFR1 and secretion of bFGF were distinctly reduced, and the inhibition efficiency of cells multiplication of WSO-38 was 48% and the inhibition efficiency of colonies of SWO-38 in soft agar was 35%. The inhibition of cells multiplication can be reversed completely by external bFGF, and the reverse efficiency was 8%.Conclusion:The synthesis of bFGF mRNA and expression of bFGF can be specifically inhibited by antisense oligonucleotide, but the inhibition can be cleared up with the addition of external bFGF. The study suggested that the bFGFantisense oligonucleotide could have good effect in inhibiting of tumor under special condition.