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1.
BMC Urol ; 23(1): 121, 2023 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-37454079

RESUMO

BACKGROUND: I kappa B kinase interacting protein, a highly conserved gene, has rarely been reported in cancer. According to previous study, IKBIP has only been shown to promote malignant progression of glioma. In other malignant tumors, few reports have examined the function of IKBIP, especially in papillary renal cell carcinoma. Therefore, the molecular profiles and clinical prognostic values of the IKBIP in papillary renal cell carcinoma remain undetermined. METHODS: Several bioinformatic platforms and Immunohistochemistry were used to clarify the expression and prognostic values of IKBIP in Papillary renal cell carcinoma. RESULTS: In this study, GEPIA and TIMER platform were used to identify mRNA expression of IKBIP in papillary renal cell carcinoma. And our results revealed that IKBIP mRNA expression was up-regulated in papillary renal cell carcinoma than in its corresponding normal tissues. In addition, high mRNA expression levels of IKBIP were correlated with age, pathological stage, pathological T stage and pathological N stage. Moreover, High IKBIP mRNA expression was negatively correlated with overall survival (OS) and disease-free survival (DFS) in patients of papillary renal cell carcinoma. Besides, Multivariate analysis indicated that IKBIP mRNA expression was an independent prognostic factor for patients of papillary renal cell carcinoma. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed IKBIP co-expressed genes were enriched in homologous recombination, DNA replication, cell cycle, Mismatch repair, Fanconi anemia pathway, P53 signaling pathway and nucleotide excision repair. And Immunohistochemical profile showed that protein expression of IKBIP was higher in papillary renal cell carcinoma than adjacent normal tissue. CONCLUSIONS: Overall, our findings reveled that IKBIP may act as a novel and potential tumor factor to accelerate papillary renal cell carcinoma progression, meanwhile, IKBIP could serve as a promising target for treating papillary renal cell carcinoma.


Assuntos
Carcinoma de Células Renais , Neoplasias Renais , Humanos , Prognóstico , Biomarcadores Tumorais/genética , RNA Mensageiro
2.
Virchows Arch ; 481(6): 847-852, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36301367

RESUMO

Metastasis is the main cause of colorectal cancer (CRC)-related death and lymph node plays a vital role in this process. Long noncoding RNAs (lncRNAs) are emerging as an important factor of biological progress in cancers. However, lncRNAs related to CRC metastasis was rarely reported.CLAN expression data of tumor tissues and normal tissues were obtained from GEPIA database and 23 paired tumor and normal samples of patients. CLAN expression of 85 patients was carried out with RNA extracted from FFPE samples and quantified with qRT-PCR. Patients' clinical features were collected from department of Pathology of the Affiliated Hospital of Southwest Medical University. Immunohistochemistry staining was used to detect the metastasis-related proteins.CLAN was highly expressed in tumor tissues. And the expression level was not correlated with age, gender, differentiation, and location of CRC patients. Also, CLAN expression did not correlated with budding, LVI, and TILs. However, CLAN expression was strongly associated with lymph node metastasis and higher TNM stage. CLAN changed the lymphatic vessel density by promoting lymphangiogenesis but CLAN did not affect the blood vessel density.CLAN was a unique lncRNA that promoted lymphangiogenesis to accelerate CRC metastasis. CLAN might play a unique role in tumor early dissemination through lymphatic vessel.


Assuntos
Proteínas Adaptadoras de Sinalização CARD , Proteínas de Ligação ao Cálcio , Neoplasias Colorretais , RNA Longo não Codificante , Humanos , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Regulação Neoplásica da Expressão Gênica , Linfangiogênese , Metástase Linfática , RNA Longo não Codificante/metabolismo , Proteínas Adaptadoras de Sinalização CARD/genética , Proteínas de Ligação ao Cálcio/genética
3.
Zhongguo Zhong Yao Za Zhi ; 43(13): 2758-2763, 2018 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-30111028

RESUMO

The expression of fibroblast growth factor 9 (FGF9) recombinant fusion protein in Carthamus tinctorius was used to identify its effect on hair regrowth and wound repair system in mice, providing a basis for C. tinctorius as a plant bioreactor, and establishing a foundation for commercial applications of FGF9 fusion protein in hair regrowth and wound repair. The identified pOTBar-oleosin-rhFGF9 plasmid was transformed into Agrobacterium tumefaciens EHA105 by freeze-thaw method, and the oleosin-rhFGF9 gene was transformed into safflower leaves by A. tumefaciens mediated method. Transgenic safflower seedlings were then obtained by tissue culture. After basta screening, transgenic T3 safflower seeds were obtained by grafting method, PCR verification and propagation. The expression of oleosin-rhFGF9 was detected by Western blot, and the content of oleosin-rhFGF9 fusion protein was 0.09% by using ELISA quantitative method. It was observed that 60 µg·L⁻¹ transgenic safflower oil had better effect on promoting NIH/3T3 cells proliferation in a certain dose-dependent manner. Sixty C57BL/6 mice were used to establish alopecia model and wound model respectively, and then were randomly divided into control group (treated with PBS or saline), negative group (treated with wild type safflower seed oil bodies, 60 g·L⁻¹), positive group (treated with FGF9, 0.054 g·L⁻¹), low dose group (treated with transgenic safflower oil bodies, 10 g·L⁻¹) and high dose group (treated with transgenic safflower oil bodies, 60 g·L⁻¹). The skin of all above-mentioned mice models were coated with soft adhesive manner every other day, 100 µL/time. After 15 days, the mice skin was cut and embedded for histological analysis. The hair regrowth experimental results showed that the hair of mice grew well, and the mice in high dose group had bushy hair, with significant effect on regeneration hair number as compared with the positive group. The healing was obvious in wound experiment, with significant healing effect in positive group, high dose group and low dose group as compared to blank control group. Furthermore, high dose group remarkably showed a better and higher healing effect than the positive group at day 5. Oleosin-rhFGF9 was successfully transformed into safflower, and T3 transgenic safflower oil bodies expressed oleosin-rhFGF9 fusion protein were obtained, with the role of promoting hair regeneration and wound repair in mice.


Assuntos
Carthamus tinctorius , Animais , Fator 9 de Crescimento de Fibroblastos , Cabelo , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Regeneração , Sementes
4.
BMC Biotechnol ; 18(1): 51, 2018 08 29.
Artigo em Inglês | MEDLINE | ID: mdl-30157831

RESUMO

BACKGROUND: Fibroblast growth factor 9 (FGF9) is a heparin-binding growth factor, secreted by both mesothelial and epithelial cells, which participates in hair follicle regeneration, wound healing, and bone development. A suitable source of recombinant human FGF9 (rhFGF9) is needed for research into potential clinical applications. We present that expression of oleosin-rhFGF9 fusion protein in safflower (Carthamus tinctorius L.) seeds stimulates hair growth and wound healing. RESULTS: The oleosin-rhFGF9 expressed in safflower seeds, in which it localizes to the surface of oil bodies. The expression of oleosin-rhFGF9 was confirmed by polyacrylamide gel electrophoresis and western blotting. According to BCA and Enzyme-linked immunosorbent assay (ELISA) assay, the results show that the expression level of oleosin-rhFGF9 was 0.14% of oil body protein. The oil body bound oleosin-rhFGF9 showed mitogenic activity towards NIH3T3 cells in a methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. The efficacy of oil body bound oleosin-rhFGF9 in promoting hair growth and wound healing was investigated in C57BL/6 mice. In a hair regeneration experiment, 50 µg/µl oil body bound oleosin-rhFGF9 was applied to the dorsal skin of mice in the resting phase of the hair growth cycle. After 15 days, thicker hair and increased number of new hairs were seen compared with controls. Furthermore, the number of new hairs was greater compared with rhFGF9-treated mice. The hair follicles of mice treated with oil body bound oleosin-rhFGF9 expressed ß-catenin more abundantly. In a wound healing experiment, dorsal skin wounds were topically treated with 50 µg/µl oil body bound oleosin-rhFGF9. Wound healing was quicker compared with mice treated with rhFGF9 and controls, especially in the earlier stages of healing. CONCLUSIONS: The oil body bound oleosin-rhFGF9 promotes both hair growth and wound healing. It appears to promote hair growth, at least in part, by up-regulating ß-catenin expression. The potential of oil body bound oleosin-rhFGF9 as an external drug can treat the alopecia and wounds or use in further clinical application.


Assuntos
Carthamus tinctorius/genética , Fator 9 de Crescimento de Fibroblastos/administração & dosagem , Fator 9 de Crescimento de Fibroblastos/genética , Cabelo/crescimento & desenvolvimento , Gotículas Lipídicas/metabolismo , Proteínas de Plantas/administração & dosagem , Proteínas de Plantas/genética , Ferimentos e Lesões/tratamento farmacológico , Animais , Carthamus tinctorius/metabolismo , Fator 9 de Crescimento de Fibroblastos/metabolismo , Expressão Gênica , Cabelo/efeitos dos fármacos , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Células NIH 3T3 , Proteínas de Plantas/metabolismo , Transporte Proteico , Proteínas Recombinantes de Fusão/administração & dosagem , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Cicatrização , Ferimentos e Lesões/genética , Ferimentos e Lesões/metabolismo , Ferimentos e Lesões/fisiopatologia , beta Catenina/genética , beta Catenina/metabolismo
5.
Int J Mol Sci ; 18(10)2017 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-29057820

RESUMO

Recombinant human fibroblast growth factor 10 (rhFGF-10) is frequently used to treat patients with skin injuries. It can also promote hair growth. However, the effective application of rhFGF-10 is limited because of its poor stability and transdermal absorption. In this study, polymerase chain reaction (PCR) and Southern blotting were used to identify transgenic safflowers carrying a gene encoding an oleosin-rhFGF-10 fusion protein. The size and structural integrity of oleosin-rhFGF-10 in oil bodies extracted from transgenic safflower seeds was characterized by polyacrylamide gel electrophoresis and western blotting. Oil body extracts containing oleosin-rhFGF-10 were topically applied to mouse skin. The absorption of oleosin-rhFGF-10 was studied by immunohistochemistry. Its efficiency in promoting wound healing and hair regeneration were evaluated in full thickness wounds and hair growth assays. We identified a safflower line that carried the transgene and expressed a 45 kDa oleosin-rhFGF-10 protein. Oil body-bound oleosin-rhFGF-10 was absorbed by the skin with higher efficiency and speed compared with prokaryotically-expressed rhFGF-10. Oleosin-rhFGF-10 also enhanced wound closure and promoted hair growth better than rhFGF-10. The application of oleosin-rhFGF-10 in oil bodies promoted its delivery through the skin, providing a basis for improved therapeutic effects in enhancing wound healing and hair growth.


Assuntos
Carthamus tinctorius/química , Portadores de Fármacos/química , Fator 10 de Crescimento de Fibroblastos/administração & dosagem , Folículo Piloso/metabolismo , Proteínas de Plantas/química , Cicatrização , Pelo Animal/efeitos dos fármacos , Pelo Animal/crescimento & desenvolvimento , Animais , Fator 10 de Crescimento de Fibroblastos/farmacocinética , Fator 10 de Crescimento de Fibroblastos/farmacologia , Folículo Piloso/efeitos dos fármacos , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL
6.
Food Chem ; 138(1): 306-14, 2013 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-23265492

RESUMO

Three sulphated polysaccharides, coded as BEMPA, BEMPB(1), BEMPB(2), were extracted from the mucilage of mud snail of Bullacta exarata and purified by DEAE-cellulose ion-exchange and size-exclusion chromatography. Structural analysis of purified polysaccharides by chemical and biochemical methods revealed BEMPA was a high (1→3,4)-linked mannose-containing polysaccharide with molecular weight of 22,977 Da. BEMPB(1), with molecular weight of 64,117 Da, was a high (1→3)-linked arabinose-containing polysaccharide. BEMPB(2) was mainly composed of (1→3,4)-linked mannose with molecular weight of 47,507Da. The comparison between sulphated polysaccharides and their desulphated products showed that sulphate substitutions of BEMPB(1) were deduced to be at the C-3 of (1→4)-linked mannose, while sulphate substitutions of BEMPA and BEMPB(2) were at C-4 of (1→3)-linked mannose. Furthermore, BEMPA exhibited highest inhibitory effects on growth of B-16 melanoma cells, and IC(50) were 31.1 µg/mL.


Assuntos
Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Caramujos/química , Animais , Estrutura Molecular , Sulfatos
7.
Eur J Pharmacol ; 608(1-3): 1-6, 2009 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-19233158

RESUMO

Ventricular tachyarrhythmias are often precipitated by physical or emotional stress, indicating a link between increased adrenergic stimulation and cardiac ion channel activity. Human ether-a-go-go related gene (hERG) potassium channels conduct the rapid component of delayed rectifier potassium current, I(kr), a crucial component for action potential repolarization. To evaluate the correlation between increased alpha(1)-adrenergic activity and the rapid component of cardiac I(kr), whole-cell patch-clamp recording was performed in isolated guinea-pig ventricular myocytes. Stimulation of alpha(1)-adrenoceptors using phenylephrine (0.1 nM-100 microM) reduced I(kr) current in a dose-dependent manner at 37 degrees C. Phenylephrine (0.1 microM) reduced I(kr) current to 66.83+/-3.16%. Chelerythrine (1 microM), a specific inhibitor of protein kinase C (PKC) completely inhibited the changes in I(kr) trigged by 0.1 microM phenylephrine. KT5720 (2.5 microM), a specific inhibitor of protein kinase A (PKA) partially inhibited the current decrease induced by 0.1 microM phenylephrine. Both chelerythrine and KT5720 drastically reduced the phenylephrine-induced effects, indicating possible involvement of PKC and PKA in the alpha(1)-adrenergic inhibition of I(kr). Our data suggest a link between I(kr) and the alpha(1)-adrenoceptor, involving activation of PKC and PKA in arrhythmogenesis.


Assuntos
Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Miócitos Cardíacos/metabolismo , Canais de Potássio Corretores do Fluxo de Internalização/metabolismo , Proteína Quinase C/metabolismo , Receptores Adrenérgicos alfa 1/metabolismo , Animais , Benzofenantridinas/farmacologia , Carbazóis/farmacologia , Proteínas Quinases Dependentes de AMP Cíclico/antagonistas & inibidores , Relação Dose-Resposta a Droga , Eletrofisiologia , Inibidores Enzimáticos/farmacologia , Cobaias , Ventrículos do Coração/citologia , Cinética , Técnicas de Patch-Clamp , Fenilefrina/farmacologia , Potássio/metabolismo , Canais de Potássio Corretores do Fluxo de Internalização/antagonistas & inibidores , Proteína Quinase C/antagonistas & inibidores , Pirróis/farmacologia
8.
Clin Exp Pharmacol Physiol ; 35(12): 1419-25, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18671725

RESUMO

1. Resveratrol, a polyphenol in red wine, has a cardioprotective effect. Resveratrol-targeting protein (RTP) has been purified using a resveratrol affinity column (RAC) and has been identified as quinone reductase type 2 (NQO2). We hypothesize that NQO2 is the target protein of resveratrol in vascular smooth muscle cells (VSMC) and that resveratrol inhibits proliferation of VSMC through its action on NQO2. In the present study, we investigated the correlation between NQO2 regulation and cell proliferation in VSMC in response to resveratrol treatment. 2. The RTP was purified using RAC and was detected with a NQO2 polyclonal antibody. The VSMC were incubated with resveratrol (1, 10 and 50 micromol/L) for 24, 48 and 72 h. Cell proliferation was detected by cell counting and bromodeoxyuridine (BrdU) assay. A lentiviral vector incorporating NQO2 short interference (si) RNA of short hairpin design was constructed and transduced into VSMC. Real-time quantitative polymerase chain reaction was used to measure NQO2 mRNA levels; NQO2 expression was determined by western blot analysis. 3. Using RAC, we extracted a 26 kDa protein from aortic smooth muscle, which was referred to as RTP-26. Proliferation of VSMC was inhibited by resveratrol in a concentration- and time-dependent manner. The mRNA and protein expression of NQO2 was also repressed by resveratrol in a concentration- and time-dependent manner. A similar pattern of inhibition was observed for cells treated with resveratrol (25 micromol/L) as for cells transduced with a lentiviral vector containing siRNA sequences against NQO2. 4. Collectively, these data indicate that the suppression of VSMC proliferation mediated by resveratrol correlates with NQO2 downregulation.


Assuntos
Regulação para Baixo/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/enzimologia , Quinona Redutases/antagonistas & inibidores , Estilbenos/farmacologia , Animais , Aorta Torácica/efeitos dos fármacos , Aorta Torácica/enzimologia , Células Cultivadas , Relação Dose-Resposta a Droga , Regulação para Baixo/fisiologia , Masculino , Quinona Redutases/biossíntese , Quinona Redutases/fisiologia , Coelhos , Ratos , Ratos Sprague-Dawley , Resveratrol , Fatores de Tempo
9.
J Appl Physiol (1985) ; 93(3): 1140-51, 2002 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-12183512

RESUMO

Despite considerable advances in medicine, the incidence of heart failure remains high in patients after myocardial infarction (MI). This study investigated the effects of engrafted early-differentiated cells (EDCs) from mouse embryonic stem cells, with or without transfection of vascular endothelial growth factor (VEGF) cDNA (phVEGF(165)), on cardiac function in postinfarcted mice. EDCs were transfected with green fluorescent protein (GFP) cDNA and transplanted into infarcted myocardium. Compared with the MI mice receiving cell-free medium, cardiac function was significantly improved in the MI mice 6 wk after transplantation of EDCs. Moreover, improvement of heart function was significantly greater in the mice implanted with EDCs overexpressing VEGF (EDCs-VEGF) than with EDCs alone. Frozen sections of infarcted myocardium with EDCs or EDCs-VEGF transplantation showed GFP-positive tissue. The area with positive immunostaining for cardiac troponin I and alpha-myosin heavy chain was larger in injured myocardium with EDCs or EDCs-VEGF transplantation than with medium injection. Transplantation of EDCs or EDCs-VEGF significantly increased the number of blood vessels in the MI area. However, the density of capillaries was significantly higher in the EDCs-VEGF animals than in the EDC mice. Double staining for GFP and connexin-43 was positive in injured myocardium with EDC transplantation. Our data demonstrate that engrafted EDCs or EDCs-VEGF regenerated cardiac tissue and significantly improved cardiac function in postinfarcted hearts. The novel EDCs-VEGF synergistic approach may have an important impact on future cell therapy for patients experiencing MI or heart failure.


Assuntos
Fatores de Crescimento Endotelial/uso terapêutico , Coração/fisiopatologia , Peptídeos e Proteínas de Sinalização Intercelular/uso terapêutico , Linfocinas/uso terapêutico , Infarto do Miocárdio/fisiopatologia , Infarto do Miocárdio/terapia , Transplante de Células-Tronco , Células-Tronco/citologia , Animais , Diferenciação Celular , Células Cultivadas , Fatores de Crescimento Endotelial/genética , Fatores de Crescimento Endotelial/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Linfocinas/genética , Linfocinas/metabolismo , Camundongos , Contração Miocárdica , Infarto do Miocárdio/patologia , Miocárdio/patologia , Células-Tronco/metabolismo , Transfecção , Fator A de Crescimento do Endotélio Vascular , Fatores de Crescimento do Endotélio Vascular
10.
Br J Pharmacol ; 135(1): 188-96, 2002 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-11786494

RESUMO

1. In the present experiments, we investigated the effects of methylecgonidine (MEG) on nitric oxide (NO) production in cultured neonatal rat cardiomyocytes. Incubation of cultured cardiomyocytes with carbachol or MEG for 48 h significantly enhanced NO production. No release was increased from 1.48+/-0.13 microM (mg protein)(-1) for control to 5.73+/-0.19 microM (mg protein)(-1) for 1 microM carbachol treated cells (P<0.001). In addition, incubation with 1 microM MEG enhanced NO production to 5.55+/-0.28 microM (mg protein)(-1). The effects of MEG on NO production were concentration-dependent. The muscarinic antagonist atropine prevented the enhancement of NO production induced by carbachol or MEG. Compared to MEG-induced NO production, cocaine was much less potent. 2. The enhancement of NO production by carbachol or MEG was even greater in cultured cardiomyocytes transfected with the M(2) cDNA. After 48-h incubation with 1 microM carbachol or 1 microM MEG, NO production was increased by 6.5 and 6.7 fold, respectively, in cardiomyocytes overexpressing M(2) receptors. Coincubation with atropine or N(G)-nitro-L-arginine methyl ester abolished the enhancement of NO production. In contrast, NO production enhanced by carbachol or MEG in M(1)- or M(3)-transfected cardiomyocytes was similar to the level in non-transfected cells. 3. Western blot analysis showed that the protein levels of M(1), M(2), and M(3) were significantly increased in cardiomyocytes transfected with the receptor cDNAs, but MEG had no effect on the expressions. It is interesting that both carbachol and MEG caused a significant increase in constitutive endothelial NO synthase (eNOS) only in M(2)-transfected cardiomyocytes, not in non-transfected, M(1)- or M(3)-transfected cells. Again, atropine blocked the MEG-produced induction of eNOS. 4. Our data demonstrate that MEG significantly enhanced NO production in cultured cardiomyocytes and that the enhancement of NO production may result from MEG stimulation of muscarinic M(2) receptors.


Assuntos
Cocaína/análogos & derivados , Cocaína/farmacologia , Miocárdio/metabolismo , Óxido Nítrico/biossíntese , Animais , Animais Recém-Nascidos , Western Blotting , Carbacol/farmacologia , Células Cultivadas , Agonistas Colinérgicos/farmacologia , AMP Cíclico/biossíntese , GMP Cíclico/biossíntese , Indução Enzimática/efeitos dos fármacos , Miocárdio/citologia , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico Sintase Tipo II , Óxido Nítrico Sintase Tipo III , Ratos , Receptores Muscarínicos/efeitos dos fármacos , Receptores Muscarínicos/genética , Receptores Muscarínicos/metabolismo , Transdução de Sinais , Transfecção
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