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1.
Acta Academiae Medicinae Sinicae ; (6): 235-244, 2023.
Artigo em Chinês | WPRIM | ID: wpr-981258

RESUMO

Objective To investigate the clinical significance of thrombospondin type 1 domain-containing 7A (THSD7A) and neural epidermal growth factor-like 1 protein (NELL1) in phospholipase A2 receptor (PLA2R)-negative membranous nephropathy (MN). Methods A total of 116 PLA2R-negative MN patients treated in Hangzhou TCM Hospital Affiliated to Zhejiang Chinese Medical University from 2014 to 2021 were enrolled in this study.Immunohistochemistry was employed to detect THSD7A and NELL1 in the renal tissue.The pathological characteristics,treatment,and prognosis were compared between positive and negative groups. Results The 116 PLA2R-negative MN patients included 23 THSD7A-positive patients and 9 NELL1-positive patients.One patient was tested positive for both proteins.The THSD7A-positive group showed higher positive rate of IgG4 (P=0.010),more obvious glomerular basement membrane (GBM) thickening (P=0.034),and higher proportion of stage Ⅱ MN and lower proportion of stage I MN (P=0.002) than the THSD7A-negative group.The NELL1-positive group had lower positive rates of C1q and IgG2 (P=0.029,P=0.001),less obvious GBM thickening (P<0.001),more extensive inflammatory cell infiltration (P=0.033),lower proportion of deposits on multi-locations (P=0.001),and lower proportion of atypical MN (P=0.010) than the NELL1-negative group.One patient with THSD7A-positive MN was diagnosed with colon cancer,while none of the NELL1-positive patients had malignancy.Survival analysis suggested that THSD7A-positive MN had worse composite remission (either complete remission or partial remission) of nephrotic syndrome than the negative group (P=0.016),whereas NELL1-positive MN exhibited better composite remission of nephrotic syndrome than the negative group (P=0.015).The MN patients only positive for NELL1 showed better composite remission of nephrotic syndrome than the MN patients only positive for THSD7A (P<0.001). Conclusions THSD7A- and NELL1-positive MN is more likely to be primary MN,and there is no significant malignancy indication.However,it might have a predictive value for the prognosis of MN.


Assuntos
Humanos , Autoanticorpos , Relevância Clínica , Neoplasias do Colo , Família de Proteínas EGF , Glomerulonefrite Membranosa/diagnóstico , Síndrome Nefrótica , Receptores da Fosfolipase A2/metabolismo , Trombospondinas/metabolismo
2.
Chinese Journal of Pediatrics ; (12): 261-265, 2023.
Artigo em Chinês | WPRIM | ID: wpr-970278

RESUMO

Objective: To summarize the genetic and clinical phenotypic characteristics of patients with early-onset myopathy, areflexia, respiratory distress and dysphagia (EMARDD) caused by multiple epidermal growth factor 10 (MEGF10) gene defect. Methods: The clinical data of 3 infants in 1 family with EMARDD caused by MEGF10 gene defect diagnosed in the Department of Neonatology, Xiamen Children's Hospital in April 2022 were analyzed retrospectively. Using "multiple epidermal growth factor 10" "myopathy" or "MEGF10" "myopathy" as the key words, and searching the relevant literature reports of CNKI, Wanfang Database and PubMed Database from the establishment of the database to September 2022. Combined with this family, the main clinical information and genotype characteristics of EMARDD patients caused by MEGF10 gene defect were summarized. Results: The proband, male, first infant of monozygotic twins, was admitted to hospital 7 days after birth "due to intermittent cyanosis with weak sucking". The infant had dysphagia accompanied with cyanosis of lips during feeding and crying after birth. Physical examination on admission revealed reduced muscle tone of the extremities, flexion of the second to fifth fingers of both hands with limited passive extension of proximal interphalangeal joints, and limited abduction of both hips. He was diagnosed as dysphagia of newborn, congenital dactyly. After admission, he was given limb and oral rehabilitation training, breathing gradually became stable and oral feeding fully allowed, and discharged along with improvement. The younger brother of the proband was admitted to the hospital at the same time, and his clinical manifestations, diagnosis and treatment process were the same as those of the proband. The elder brother of the proband died at the age of 8 months due to the delayed growth and development, severe malnutrition, hypotonia, single palmoclal crease and weak crying. A whole exon sequencing of the family was done, and found that the 3 children were all compound heterozygous variations at the same site of MEGF10 gene, with 2 splicing variants (c.218+1G>A, c.2362+1G>A), which came from the father and mother respectively, and the new variation was consistent with the autosomal recessive inheritance model. Three children were finally diagnosed as EMARDD caused by MEGF10 gene defect. There are 0 Chinese literature and 18 English literature that met the search conditions. Totally 17 families including 28 patients were reported. There were 31 EMARDD patients including 3 infants from this family. Among them, there were 13 males and 18 females. The reported age of onset ranged from 0 to 61 years. Except for 5 patients with incomplete clinical data, 26 patients were included in the analysis of phenotypic and genotypic characteristics. The clinical features were mainly dyspnea (25 cases), scoliosis (22 cases), feeding difficulties (21 cases), myasthenia (20 cases), and other features including areflexia (16 cases) and cleft palate or high palatal arch(15 cases). Muscle biopsy showed non-specific changes, with histological characteristics ranging from slight muscle fiber size variation to minicores change which was seen in all 5 patients with at least 1 missense mutation of allele. In addition, the adult onset was found in patients with at least 1 missense variant of MEGF10 gene. Conclusions: MEGF10 gene defect related EMARDD can occur in the neonatal period, and the main clinical features are muscle weakness, breathing and feeding difficulties. Patients with myopathy who have at least 1 missense mutation and muscle biopsy indicating minicores change may be relatively mild.


Assuntos
Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Cianose , Transtornos de Deglutição , Família de Proteínas EGF , Hipotonia Muscular , Debilidade Muscular , Doenças Musculares/genética , Estudos Retrospectivos
3.
Chinese Journal of Burns ; (6): 305-311, 2016.
Artigo em Chinês | WPRIM | ID: wpr-327342

RESUMO

<p><b>OBJECTIVE</b>To investigate the suitable transfection condition of human epidermal cells (hECs) with human epidermal growth factor (EGF) gene by adenovirus vector (Ad-hEGF) and its effects on the biological characteristics of hECs.</p><p><b>METHODS</b>hECs were isolated from deprecated human fresh prepuce tissue of circumcision by enzyme digestion method and then sub-cultured. hECs of the third passage were used in the following experiments. (1) Cells were divided into non-transfection group and 5, 20, 50, 100, 150, and 200 fold transfection groups according to the random number table (the same grouping method below), with 3 wells in each group. Cells in non-transfection group were not transfected with Ad-hEGF gene, while cells in the latter six groups were transfected with Ad-hEGF gene in multiplicities of infection (MOI) of 5, 20, 50, 100, 150, and 200 respectively. The morphology of the cells was observed with inverted phase contrast microscope, and expression of green fluorescent protein of the cells was observed with inverted fluorescence microscope at transfection hour (TH) 24, 48, and 72. (2) Another three batches of cells were collected, grouped, and treated as above, respectively. Then the transfection rate of Ad-hEGF gene was detected by flow cytometer (n=3), the mass concentration of EGF in culture supernatant of cells was detected by enzyme-linked immunosorbent assay (n=6), and the proliferation activity of cells was detected by cell counting kit 8 (CCK8) and microplate reader (n=6) at TH 24, 48, and 72, respectively. (3) Cells were collected and divided into non-transfection group and transfection group, with 6 wells in each group. Cells in non-transfection group were cultured with culture supernatant of cells without transfection, while cells in transfection group were cultured with culture supernatant of cells which were transfected with Ad-hEGF gene in the optimum MOI (50). CCK8 and microplate reader were used to measure the biological activity of EGF secreted by cells on culture day 1, 3, and 5. (4) Cells were collected and divided into non-transfection group and transfection group, with 12 wells in each group. Cells in non-transfection group were not transfected with Ad-hEGF gene, while cells in transfection group were transfected with Ad-hEGF gene in the optimum MOI (50). The expression levels of cytokeratin 14 (CK14) and CK19 of cells were measured by immunofluorescence staining at TH 24. (5) Cells were collected, grouped, and treated as in (4), with 6 wells in each group. At post scratch hour (PSH) 0 (immediately after scratch), 12, 24, and 48, the migration distance of cells was observed and measured with inverted phase contrast microscope. Data were processed with analysis of variance of factorial design, analysis of variance for repeated measurement, and LSD test.</p><p><b>RESULTS</b>(1) At TH 24 and 48, morphology of cells in each transfection group and non-transfection group were similar. Compared with that in non-transfection group, the cell debris increased significantly in 200 fold transfection group at TH 72. At TH 24, 48, and 72, the expression of green fluorescent protein was not seen in cells of non-transfection group, whereas it increased in cells of transfection group over transfection time. (2) The transfection rate of Ad-hEGF gene of cells in each transfection group increased gradually over transfection time. At TH 72, the transfection rates of Ad-hEGF gene of cells in 50-200 fold transfection groups were all above 90%, while the transfection rates of Ad-hEGF gene of cells in non-transfection group, 5, and 20 fold transfection groups were (0.51±0.20)%, (62.44±6.23)%, and (75.00±5.43)% respectively, which were obviously lower than the rate in 50 fold transfection group [(93.12±2.55)%, with P values below 0.01]. The mass concentration of EGF in culture supernatant of cells in each transfection group increased gradually over transfection time. At TH 72, the mass concentration of EGF in culture supernatant of cells in 50 fold transfection group was obviously higher than that in each of the other groups (with P values below 0.01). The proliferation activity of cells in each group at TH 24 and 48 was similar (with P values above 0.05). At TH 72, the proliferation activity of cells in 200 fold transfection group was obviously lower than that in other groups (with P values below 0.05). (3) On culture day 1, the biological activity of EGF secreted by cells in two groups was similar (P>0.05). On culture day 3 and 5, the biological activity of EGF secreted by cells in transfection group were obviously higher than that in non-transfection group (with P values below 0.01). (4) At TH 24, the expression levels of CK14 and CK19 of cells in transfection group were higher than those in non-transfection group. (5) The width of scratch in two groups was nearly the same at PSH 0. At PSH 12-48, the migration distance of cells in transfection group was obviously longer than that in non-transfection group (with P values below 0.01).</p><p><b>CONCLUSIONS</b>The suitable range of MOI of hECs transfected with Ad-hEGF gene is 50-150, and 50 is the optimum. hECs transfected with Ad-hEGF gene with MOI 50 can effectively express the EGF gene and keep its good abilities of proliferation, differentiation, and migration, as well.</p>


Assuntos
Humanos , Masculino , Adenoviridae , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Família de Proteínas EGF , Genética , Metabolismo , Epiderme , Biologia Celular , Vetores Genéticos , Queratinas , Metabolismo , Transfecção
4.
Journal of Central South University(Medical Sciences) ; (12): 507-512, 2016.
Artigo em Chinês | WPRIM | ID: wpr-815006

RESUMO

OBJECTIVE@#To investigate androgen receptor (AR) expression and the effect of epidermal growth factor (EGF) and testosterone on AR expression level.
@*METHODS@#EGF or different concentrations of testosterone were incubated with the primary urethral plate fibroblasts from patients with hypospadias. The levels of AR expression in the fibroblasts were detected by immunocytochemical assays and graphical analysis.
@*RESULTS@#There was no significant difference in AR activation under physiological concentrations (3×10(-8) mol/L) of testosterone between the control and the distal hypospadias group (P>0.05). However, there was a significant decrease in AR activation in the proximal hypospadias group compared to that in the control group (Pdistal hypospadias group>proximal hypospadias group, P<0.001). AR activation level in the group of proximal hypospadias was improved most obviously when EGF and physiological concentration of testosterone were employed in the urethral plate fibroblasts from hypospadias patients (P<0.001), and it was improved more in the distal hypospadias group than that in the control group (P=0.02).
@*CONCLUSION@#AR expression and activation in the urethral plate fibroblasts from hypospadias patients are abnormal. EGF can be used to improve AR activation in fibroblasts from different types of hypospadias, especially in the proximal type.


Assuntos
Humanos , Masculino , Células Cultivadas , Família de Proteínas EGF , Metabolismo , Fibroblastos , Metabolismo , Hipospadia , Metabolismo , Receptores Androgênicos , Metabolismo , Testosterona , Farmacologia
5.
Journal of Southern Medical University ; (12): 221-225, 2016.
Artigo em Chinês | WPRIM | ID: wpr-273784

RESUMO

<p><b>OBJECTIVE</b>To investigate effects of retinol on the expressions of epidermal growth factor (EGF), stem cell factor (SCF), colony-stimulating factor 1 (CSF1) and leukemia inhibitory factor (LIF) in cultured human umbilical-derived mesenchymal stem cells (UCMSCs).</p><p><b>METHODS</b>Human UCMSCs were isolated from human umbilical cord and identified for immunophenotypes. The cells were then cultured in DMEM/F12 media supplemented with 12% fetal bovine serum (FBS), 12% FBS+1 µmol/L retinol, 15% knockout serum replacement (KSR) and 15% KSR+ 1 µmol/L retinol. The expressions of the cytokines EGF, SCF, CSF1 and LIF in the cells were detected using RT-PCR and ELISA.</p><p><b>RESULTS</b>The isolated cells exhibited characteristic immunophenotypes of human UCMSCs and expressed EGF, CSF1 and SCF at both mRNA and protein levels but not LIF protein. Retinol (1 µmol/L) significantly promoted the expressions of SCF and CSF1 at both mRNA and protein levels but did not result in changes of EGF and LIF expressions in human UCMSCs.</p><p><b>CONCLUSION</b>Retinol at the concentration of 1 µmol/L can promote expression of SCF and CSF1 in human UCMSCs in vitro.</p>


Assuntos
Humanos , Diferenciação Celular , Células Cultivadas , Família de Proteínas EGF , Metabolismo , Imunofenotipagem , Fator Inibidor de Leucemia , Metabolismo , Fator Estimulador de Colônias de Macrófagos , Metabolismo , Células-Tronco Mesenquimais , Metabolismo , Fator de Células-Tronco , Metabolismo , Cordão Umbilical , Biologia Celular , Vitamina A , Farmacologia
6.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 327-332, 2015.
Artigo em Inglês | WPRIM | ID: wpr-331065

RESUMO

Previous studies suggested an association between the EGF +61 A>G polymorphism and susceptibility to gastric cancer, but the results have been inconsistent. To draw a more precise risk estimation of the association, we performed a meta-analysis of published studies. PubMed, EMBASE, Google Scholar and the Chinese Wanfang databases were systematically searched to identify relevant studies. There were 7 studies involving 1992 cases of gastric cancer and 3202 controls in this meta-analysis. Our study showed that, overall, the EGF +61 A>G polymorphism was significantly associated with the increased risk of gastric cancer in allele model (G vs. A: OR=1.18, 95% CI=1.00-1.39), dominant model (GG + GA vs. AA: OR=1.28, 95% CI=1.05-1.55), homozygous model (GG vs. AA: OR=1.31, 95% CI=1.06-1.63) and heterozygous model (GA vs. AA: OR=1.25, 95% CI=1.01-1.53). The stratified analysis by ethnicity revealed a significant association between EGF +61 A>G polymorphism and gastric cancer risks in Asians. This meta-analysis indicates that EGF +61 A>G polymorphism may increase the risk of gastric cancer, especially in Asians. Large-sized, well-designed studies involving different ethnic groups should be conducted to confirm this association.


Assuntos
Humanos , Povo Asiático , Genética , Família de Proteínas EGF , Genética , Fator de Crescimento Epidérmico , Genética , Estudos de Associação Genética , Predisposição Genética para Doença , Polimorfismo de Nucleotídeo Único , Análise de Regressão , Neoplasias Gástricas , Genética
7.
West China Journal of Stomatology ; (6): 441-445, 2014.
Artigo em Chinês | WPRIM | ID: wpr-231830

RESUMO

<p><b>OBJECTIVE</b>This study aims to investigate the expression levels of serum and urinary vascular endothelial growth factor-A (VEGF-A) and epidermal growth factor-like domain 7 (EGFL7) in proliferating infantile hemangioma patients under propranolol treatment.</p><p><b>METHODS</b>Propranolol (0.5-2 mg x kg(-1)) was orally administered to 30 infants every day for 4-8 months. The Achauer method was used to measure the tumor radius and thus evaluate the clinical curative effects of the treatment. Enzyme-linked immunosorbent assay was used to measure the serum and urinary concentrations of VEGF-A and EGFL7 at 0, 4, and 12 weeks after the treatment.</p><p><b>RESULTS</b>The treatment response was excellent in 2 patients, good in 11, moderate in 14, and poor in 3. Serum VEGF-A (335.692 pg x mL(-1) ± 136.146 pg x mL(-1)) was high before the treatment and then significantly decreased after 4 weeks (264.853 pg x mL(-1) ± 122.120 pg x mL(-1)) and 12 weeks (211.345 pg x mL(-1) ± 104.035 pg x mL(-1)) of treatment (P < 0.05). Urinary VEGF-A (76.234 pg x mL(-1) ± 24.169 pg x mL(-1)) was high before the treatment and then significantly decreased after four weeks (56.454 pg x mL(-1) ± l6.111 pg x mL(-1)) and twelve weeks (34.728 pg x mL(-1)) ± 12.656 pg x mL(-1)) of treatment (P < 0.05). Serum and urinary EGFL7 also decreased after the treatment, showing a positive relationship with VEGF-A.</p><p><b>CONCLUSION</b>Propranolol can be safely and effectively used to treat proliferating infantile hemangiomas. This treatment can reduce the peripheral serum and urinary concentrations of VEGF-A and EGFL7 in affected children.</p>


Assuntos
Humanos , Lactente , Família de Proteínas EGF , Ensaio de Imunoadsorção Enzimática , Hemangioma , Propranolol , Fator A de Crescimento do Endotélio Vascular
8.
Chinese Journal of Otorhinolaryngology Head and Neck Surgery ; (12): 643-648, 2014.
Artigo em Chinês | WPRIM | ID: wpr-233831

RESUMO

<p><b>OBJECTIVE</b>To explore the effects of epidermal growth factor-like domain 7 (EGFL7) gene silencing on the proliferation and invasion ablity of laryngeal carcinoma cells.</p><p><b>METHODS</b>A lentiviral vector expressing EGFL7 shRNA was constructed and transfected into human laryngeal carcinoma Hep-2 cells. The expressions of EGFL7 mRNA and protein were detected by Real-time PCR and Western blot, respectively. Cell proliferation was evaluated by CCK-8 assay, cell cycle and apoptosis were tested by flow cytometry, and cell invasion was detected by transwell invasion assay.</p><p><b>RESULTS</b>The relative expression level s of EGFL7 mRNA and protein in EGFL7-SuRNA group were svgnificantly lower than control group (P < 0.001). Western blot analysis proved that the relative expression of EGFL7 protein in NC group, Lenti-NC group and Lenti-EGFL7 group was (0.39 ± 0.12),(0.36 ± 0.14) and (0.07 ± 0.04), respectively. EGFL7 expression in Lenri-EGFL7 group was significantly inhibited than NC group (P < 0.001), which confirmed that the recombinant lentivirus was successfully transfected into Hep-2 cells. The proliferation of Hep-2 cells was significantly inhibited after transfection (P < 0.01). Compared with the NC group and Lenti-NC group, the proportion of cells in S phase was significantly increased in Lenti-EGFL7 group (P < 0.01), and the proportion in G1 phase was significantly decreased (P < 0.05). Cell apoptosis assay showed that the apoptotic rate in Lenti-EGFL7 group (66.2 ± 1.28) % was significantly increased in NC group (6.09 ± 3.28) % and Lenti-NC group (9.86 ± 2.13) %. In Transwell invision assay, the mean number of cells coming through the Metrigel in Lenti-EGFL7 group was significantly decreased than in the NC group and Lenti-NC group (P < 0.01).</p><p><b>CONCLUSION</b>The proliferation and invasion ablity of laryngeal carcinoma Hep-2 cells can be inhibited by siRNA mediated EGFL7 gene silencing.</p>


Assuntos
Humanos , Apoptose , Ciclo Celular , Divisão Celular , Linhagem Celular Tumoral , Proliferação de Células , Família de Proteínas EGF , Inativação Gênica , Vetores Genéticos , Células Hep G2 , Neoplasias Laríngeas , Metabolismo , Virologia , Lentivirus , RNA Mensageiro , RNA Interferente Pequeno , Transfecção
9.
International Journal of Oral Science ; (4): 14-20, 2013.
Artigo em Inglês | WPRIM | ID: wpr-358201

RESUMO

In the present study, we investigate the expression profile of the epidermal growth factor receptor family, which comprises EGFR/ErbB1, HER2/ErbB2, HER3/ErbB3 and HER4/ErbB4 in oral leukoplakia (LP). The expression of four epidermal growth factor receptor (EGFR) family genes and their ligands were measured in LP tissues from 14 patients and compared with levels in 10 patients with oral lichen planus (OLP) and normal oral mucosa (NOM) from 14 healthy donors by real-time polymerase chain reaction (PCR) and immunohistochemistry. Synchronous mRNA coexpression of ErbB1, ErbB2, ErbB3 and ErbB4 was detected in LP lesions. Out of the receptors, only ErbB4 mRNA and protein was more highly expressed in LP compared with NOM tissues. These were strongly expressed by epithelial keratinocytes in LP lesions, as shown by immunohistochemistry. Regarding the ligands, the mRNA of Neuregulin2 and 4 were more highly expressed in OLP compared with NOM tissues. Therefore, enhanced ErbB4 on the keratinocytes and synchronous modulation of EGFR family genes may contribute to the pathogenesis and carcinogenesis of LP.


Assuntos
Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Anfirregulina , Betacelulina , Família de Proteínas EGF , Fator de Crescimento Epidérmico , Metabolismo , Epirregulina , Perfilação da Expressão Gênica , Glicoproteínas , Metabolismo , Heparina , Metabolismo , Fator de Crescimento Semelhante a EGF de Ligação à Heparina , Peptídeos e Proteínas de Sinalização Intercelular , Metabolismo , Queratinócitos , Metabolismo , Leucoplasia Oral , Metabolismo , Líquen Plano Bucal , Metabolismo , Ligantes , Mucosa Bucal , Metabolismo , Fatores de Crescimento Neural , Neurregulinas , Metabolismo , RNA Mensageiro , Metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Receptores ErbB , Metabolismo , Receptor ErbB-2 , Metabolismo , Receptor ErbB-3 , Metabolismo , Receptor ErbB-4 , Receptores de Superfície Celular , Metabolismo , Fator de Crescimento Transformador alfa , Metabolismo , Regulação para Cima , Fisiologia
10.
Acta Pharmaceutica Sinica ; (12): 809-813, 2013.
Artigo em Chinês | WPRIM | ID: wpr-259546

RESUMO

Psoriasis is a chronic inflammatory disease related to genome-wide and surroundings, it is important to develop a suitable animal model to research psoriasis pathogenesis and evolve pharmacotherapeutics. With the development of transgenetic technology in the past few years, psoriasis virulence gene animal model become a hotspot. Research of animal model of human psoriasis genes is reviewed in the paper.


Assuntos
Animais , Humanos , Aminoquinolinas , Toxicidade , Anfirregulina , Modelos Animais de Doenças , Família de Proteínas EGF , Genética , Metabolismo , Queratina-14 , Genética , Metabolismo , Queratina-5 , Genética , Metabolismo , Queratinócitos , Metabolismo , Glicoproteínas de Membrana , Camundongos Transgênicos , Psoríase , Genética , Metabolismo , Receptor TIE-2 , Genética , Metabolismo , Fator de Transcrição STAT3 , Genética , Metabolismo , Receptor 7 Toll-Like , Fator de Crescimento Transformador beta1 , Genética , Metabolismo
11.
West China Journal of Stomatology ; (6): 364-367, 2012.
Artigo em Chinês | WPRIM | ID: wpr-322382

RESUMO

<p><b>OBJECTIVE</b>The purpose of this study was to investigate the regulatory role of tyrosine kinase 2 with immunoglobulin-like and epidermal growth factor homology domains (Tie2) on apoptosis and proliferation in the endothelial cells.</p><p><b>METHODS</b>RNA interference (RNAi) technique was used to silence Tie2 gene expression by transfecting an expression vector containing short hairpin RNA(shRNA) for Tie2 into human umbilical vein endothelial cells (HUVECs). Real time quantitation reverse transcriptase polymerase chain reaction (QRT-PCR) and Western blot were used to monitor Tie2 mRNA, as well as protein expression. The proliferation of HUVECs was examined by methyl thiazolyl tetrazolium (MTT), and the apoptosis was detected under microscope. HUVECs transfected with pGenesil-hk was negative control, and HUVECs transfected with nothing was empty control.</p><p><b>RESULTS</b>Tie2 mRNA expression was down-regulated 24 h and 48 h after transfection, and Tie2 protein expression was significantly down-regulated at 24 h and 48 h (P< 0.05), especially 48 h after transfection. The apoptosis rate was conspicuously higher in experimental group than in negative control and empty control group after 48 h (P<0.05). The growth monitoring showed that proliferation was also markedly inhibited in experimental group (P<0.05) compared with two control groups.</p><p><b>CONCLUSION</b>Down-regulated expression of Tie2 by RNAi can promotes apoptosis of HUVECs and has an anti-proliferation activity effect on them.</p>


Assuntos
Humanos , Apoptose , Linhagem Celular Tumoral , Proliferação de Células , Regulação para Baixo , Família de Proteínas EGF , Células Endoteliais da Veia Umbilical Humana , Imunoglobulinas , Interferência de RNA , RNA Mensageiro , RNA Interferente Pequeno , TYK2 Quinase , Transfecção
12.
West China Journal of Stomatology ; (6): 653-656, 2009.
Artigo em Chinês | WPRIM | ID: wpr-242927

RESUMO

<p><b>OBJECTIVE</b>To study the cultural method and identification of human umbilical vein endothelial cells (HUVECs), and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains(Tie-2) in HUVECs.</p><p><b>METHODS</b>HUVECs were isolated from umbilical veins by the technique of irrigative digestion, and were cultivated in plates. The cells were identified by VIII monoclonal antibody. Tie-2 mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and SABC immunocytochemistry.</p><p><b>RESULTS</b>HUVECs could adhere to the plates completely after 24 hours, and confluence a monolayer 4-5 days later. The band of Tie-2 mRNA was obviously and the expression of Tie-2 protein was strongly positive by immunocytochemistry in HUVECs. The positive rate was over 85%.</p><p><b>CONCLUSION</b>Highly purified endothelial cells were isolated. And there were overexpression of Tie-2 in HUVECs.</p>


Assuntos
Humanos , Células Cultivadas , Família de Proteínas EGF , Células Endoteliais , Células Endoteliais da Veia Umbilical Humana , Imunoglobulinas , TYK2 Quinase , Veias Umbilicais
13.
West China Journal of Stomatology ; (6): 36-39, 2008.
Artigo em Chinês | WPRIM | ID: wpr-296722

RESUMO

<p><b>OBJECTIVE</b>To examine the change of epidermal growth factor (EGF) concentration in saliva of recurrent aphthous ulcer (RAU) patients during the ulcerous and interval period and epidermal growth factor receptor (EGFR) in ulcer biopsy samples.</p><p><b>METHODS</b>ECF data of the samples, which were 27 saliva samples from RAU gained not only in the ulcerous period but also in interval period and 33 ones from normal persons, were acquired through enzyme linked immunosorhent assay (ELISA) and EGF standard curve. ECFR-RNA date of RAU biopsies, which were 31 biopsy samples from RAU got during the ulcerous period and 35 ones from normal persons, were surveyed by QF-RT-PCR. All RAU samples were obtained under the same level, which were the whole patients were minor aphthous ulcers and their ulcers occurred not over the first four days. All patients and normal persons were selected seriously under the rule of physical situations without any other diseases and histories of using medicines.</p><p><b>RESULTS</b>The EGF concentration of saliva in RAU group at ulcer occurrence was higher than that in the interval period and the normal control with a significant test (F = 3.24, P < 0.05). The EGF concentration of saliva in RAU group during the interval period was lower than that in the control, which was significant on statistics (t = 2.73, P < 0.05). The EGFR-RNA in RAU group at ulcer occurrence was higher than the normal control with a significant test (t = 3.15, P < 0.05).</p><p><b>CONCLUSION</b>It was suggested that the ulcer occasion of RAU patients could be related with the decreasing of EGF in saliva during interval period, and that the ulcer sell-cure of RAU patients would be contributed to</p>


Assuntos
Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Biópsia , Família de Proteínas EGF , Receptores ErbB , Saliva , Estomatite Aftosa
14.
Chinese Journal of Oncology ; (12): 521-523, 2005.
Artigo em Chinês | WPRIM | ID: wpr-358582

RESUMO

<p><b>OBJECTIVE</b>To investigate the anti-angiogenic effect of amphiregulin (AR) antisense RNA expression in breast cancer.</p><p><b>METHODS</b>Human AR cDNA antisense plasmid was transfected into NS2T2A1 cells (a human breast cancer cell line). Two selected clones expressed AR antisense RNA (AR AS1 and AR AS3 cell lines) in which AR protein expression was reduced. Control cell line NS2T2A1 V was obtained by empty vector transfection. These cells were injected subcutaneously into nude mice. The effects of conditioned media on proliferation of human microvascular endothelial cells (HMEC) were evaluated and VEGF secreted by the cells was measured by ELISA method. In tumor tissues, VEGF expression levels were measured by quantitative RT-PCR, and CD31-immunostaining was used for intra-tumoral vascular quantification.</p><p><b>RESULTS</b>The proliferation index of HMEC cells grown in conditioned media with AR AS1 and AR AS3 was significantly reduced in comparison with that of control cells, accompanied by a decreased VEGF secretion. In tumors derived from AR AS1 and AR AS3 cells, intra-tumoral vascularization was reduced to about 50% of that derived from control cell line, accompanied with a decrease of VEGF expression.</p><p><b>CONCLUSION</b>Amphiregulin antisense RNA expression inhibits efficiently the angiogenesis in breast cancer, suggesting this growth factor could represent a novel therapeutic target in breast cancer.</p>


Assuntos
Animais , Humanos , Camundongos , Adenocarcinoma , Patologia , Anfirregulina , Inibidores da Angiogênese , Genética , Neoplasias da Mama , Patologia , Família de Proteínas EGF , Glicoproteínas , Genética , Peptídeos e Proteínas de Sinalização Intercelular , Genética , Camundongos Nus , Neovascularização Patológica , RNA Antissenso , Genética , Fatores de Crescimento do Endotélio Vascular
15.
Chinese Journal of Stomatology ; (12): 294-296, 2002.
Artigo em Inglês | WPRIM | ID: wpr-244813

RESUMO

<p><b>OBJECTIVE</b>To detect the expression and distribution of epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) in periodontal tissues, and analyze the role of EGF in orthodontic tooth movement.</p><p><b>METHODS</b>According to Kings methods, 40 g mesial force was applied to pull the left maxillary first molar in the rat. Using immunohistochemical method (HI-SABC method) to localize and examine the expression of EGF and EGFR in decalcified alveodental connective tissues at 24 hours and 168 hours of tooth movement.</p><p><b>RESULTS</b>EGF and EGFR were stained at some of periodontal ligament of furcation and radical regions in control group. These expressions of EGF and EGFR increased in periodontal tissues (P < 0.01), with the expressions at 168 hours higher than those at 24 hours (P < 0.01). And levels of EGF and EGFR at tension side were higher than those at pressure side at the same time (P < 0.01).</p><p><b>CONCLUSIONS</b>Epidermal growth factor participated in the tissues remodeling during orthodontic tooth movement and especially played a more important role in orthodontic bone formation.</p>


Assuntos
Animais , Ratos , Família de Proteínas EGF , Dente Molar , Ligamento Periodontal , Metabolismo , Periodonto , Metabolismo , Receptores ErbB , Técnicas de Movimentação Dentária
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