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1.
Zhongguo Zhong Yao Za Zhi ; 44(16): 3520-3525, 2019 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-31602917

RESUMO

The effect of triptolide( TP) on VEGFA,SDF-1,CXCR4 pathway were investigated in vitro to explore the mechanism in improving platelet activation in patients with ankylosing spondylitis( AS). Peripheral blood mononuclear cells( PBMC) were used for the experiment and divided into 4 groups: normal group( NC),model group( MC),triptolide group( TP),and AMD3100 group. The optimal concentration of TP was measured by the MTT method. The expressions of TNF-α,IL-1ß,IL-4,IL-10,VEGFA and VEGFR were detected by ELISA. The expressions of SDF-1,CXCR4 and VEGFA were detected by real-time quantitative PCR( RT-qPCR).The expressions of SDF-1,CXCR4,VEGFA and VEGFR were detected by Western blot. The expression levels of CD62 p,CD40 L and PDGFA were detected by immunofluorescence. MTT results showed that medium-dose TP had the strongest inhibitory effect on cells at24 h. The results of ELISA and PCR showed that TP inhibited mRNA expressions of IL-1ß,TNF-α,VEGFA,VEGFR and SDF-1,CXCR4 and VEGFA. The results of Western blot indicated that TP inhibited SDF-1,CXCR4 and VEGFA,VEGFR protein expressions; immunofluorescence results indicate that TP can inhibit the expressions of CD62 p,CD40 L,PDGFA. TP may regulate platelet activation by down-regulating SDF-1,CXCR4,VEGFA and VEGFR mRNA expressions,thereby down-regulating IL-1ß and TNF-αexpressions,and up-regulating the expressions of IL-4 and IL-10 cytokines.


Assuntos
Diterpenos/farmacologia , Leucócitos Mononucleares/efeitos dos fármacos , Fenantrenos/farmacologia , Ativação Plaquetária , Espondilite Anquilosante , Células Cultivadas , Quimiocina CXCL12/metabolismo , Citocinas/metabolismo , Compostos de Epóxi/farmacologia , Compostos Heterocíclicos/farmacologia , Humanos , Receptores CXCR4/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo
2.
Cell Biochem Funct ; 37(7): 504-515, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31368195

RESUMO

The treatment of neural deficiency after cerebral infarction is challenging, with limited therapeutic options. The transplantation of mesenchymal stem cells (MSCs) to the ischemic penumbra is a potential therapeutic approach. In the present study, a cerebral infarction model was generated by performing middle cerebral artery occlusion (MCAO) in SD rats. The expression of CXCR4 increased, and the number of MSCs migrating to the peri-infarct area was higher in rats transplanted with preconditioned MSCs than in rats transplanted with untreated MSCs. The rate of apoptosis, as evaluated by TUNEL staining and immunoblotting assays, was reduced in rats receiving preconditioned MSCs. A significant amelioration of neural regeneration and improved neurological function were observed in rats injected with preconditioned MSCs compared with those injected with untreated MSCs. However, the application of an siRNA targeting CXCL12 significantly inhibited the protective role of preconditioned MSCs against apoptosis and promoted the migration of MSCs to the ischemic area, leading to impaired neuronal regeneration and limited recovery of neuronal function. Hypoxic preconditioning of MSCs prior to transplantation suppressed apoptosis and increased their migration abilities, leading to the promotion of neuronal regeneration and improvement in neural function after transplantation. This preconditioning strategy may be considered as a potential approach for the modification of MSCs prior to cell transplantation therapy in patients with cerebral infarction. SIGNIFICANCE OF THE STUDY: We found that hypoxic preconditioning of MSCs improved their ability to promote neuronal regeneration and the recovery of neuronal function. Moreover, we showed that CXCR4 inhibited apoptosis, improved cell homing, and promoted neuronal differentiation, without influencing angiogenesis. Our study provides a relatively safe preconditioning method for potential use for cell transplantation therapy in ischemic cerebral infarction. The results presented here will facilitate the development of novel strategies and techniques to improve the tolerance and migration ability of transplanted cells for the treatment of cerebral infarction sequelae.


Assuntos
Infarto Cerebral/metabolismo , Quimiocina CXCL12/metabolismo , Modelos Animais de Doenças , Hipóxia , Células-Tronco Mesenquimais/metabolismo , Receptores CXCR4/metabolismo , Animais , Apoptose/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular , Infarto Cerebral/terapia , Quimiocina CXCL12/antagonistas & inibidores , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/efeitos dos fármacos , Neurônios , RNA Interferente Pequeno/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores CXCR4/antagonistas & inibidores , Transdução de Sinais/efeitos dos fármacos
3.
Cancer Sci ; 110(10): 3197-3203, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31361379

RESUMO

Intrahepatic cholangiocarcinoma is a rare malignant biliary neoplasm that causes a poor prognosis even after curative hepatectomy. Liver metastasis is the major recurrence pattern of intrahepatic cholangiocarcinoma; therefore, the prevention of liver metastasis is a desirable objective. The aim of this study is to identify gene(s) related to liver metastasis of intrahepatic cholangiocarcinoma and to examine the inhibitory effects on metastasis of intrahepatic cholangiocarcinoma by controlling such gene(s). We collected 3 pairs of intrahepatic cholangiocarcinoma frozen samples, and 36 pairs (primary and metastatic lesions) of intrahepatic cholangiocarcinoma formalin-fixed paraffin-embedded samples, from patients who underwent surgical resection at hospitals related to the Kyushu Study Group of Liver Surgery between 2002 and 2016. We carried out cDNA microarray analyses and immunohistochemistry to identify candidate genes, and evaluated one of them as a therapeutic target using human cholangiocarcinoma cell lines. We identified 4 genes related to liver metastasis using cDNA microarray, and found that CXCL12 was the only gene whose expression was significantly higher in liver metastasis than in primary intrahepatic cholangiocarcinoma by immunohistochemistry (P = .003). In prognosis, patients in the high CXCL12 group showed a significantly poor prognosis in disease-free (P < .0001) and overall survival (P = .0004). By knockdown of CXCL12, we could significantly suppress the invasive and migratory capabilities of 2 human cholangiocarcinoma cell lines. Therefore, CXCL12 might be associated with metastasis and poor prognosis in intrahepatic cholangiocarcinoma.


Assuntos
Neoplasias dos Ductos Biliares/patologia , Quimiocina CXCL12/genética , Quimiocina CXCL12/metabolismo , Colangiocarcinoma/patologia , Neoplasias Hepáticas/secundário , Idoso , Neoplasias dos Ductos Biliares/genética , Neoplasias dos Ductos Biliares/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Colangiocarcinoma/genética , Colangiocarcinoma/metabolismo , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Prognóstico , Regulação para Cima
4.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 33(6): 689-697, 2019 Jun 15.
Artigo em Chinês | MEDLINE | ID: mdl-31197995

RESUMO

Objective: To observe the change of stromal cell-derived factor 1α/cysteine X cysteine receptor 4 (SDF-1α/CXCR4) signaling pathway during the process of axial stress stimulation promoting bone regeneration, and to further explore its mechanism. Methods: A total of 72 male New Zealand white rabbits were selected to prepare the single cortical bone defect in diameter of 8 mm at the proximal end of the right tibia that repaired with deproteinized cancellous bone. All models were randomly divided into 3 groups ( n=24). Group A was treated with intraperitoneally injection of PBS; Group B was treated with stress stimulation and intraperitoneally injection of PBS; Group C was treated with stress stimulation and intraperitoneally injection of AMD3100 solution. The X-ray films were taken and Lane-Sandhu scores of bone healing were scored at 2, 4, 8, and 12 weeks after operation, while specimens were harvested for HE staining, immunohistochemical staining of vascular endothelial growth factor (VEGF) and CXCR4, and Western blot (SDF-1α and CXCR4). The bone healing area was scanned by Micro-CT at 12 weeks after operation, and the volume and density of new bone were calculated. Results: X-ray film showed that the Lane-Sandhu scores of bone healing in group B were significantly higher than those in groups A and C at 4, 8, and 12 weeks after operation ( P<0.05). Micro-CT scan showed that the bone defect was repaired in group B and the pulp cavity was re-passed at 12 weeks after operation. The volume and density of new bone were higher in group B than in groups A and C ( P<0.05). HE staining showed that the new bone growth in bone defect area and the degradation of scaffolds were faster in group B than in groups A and C after 4 weeks. The immunohistochemical staining showed that the expressions of VEGF and CXCR4 in 3 groups reached the peak at 4 weeks, and group B was higher than groups A and C ( P<0.05). Western blot analysis showed that the expressions of SDF-1α and CXCR4 in group B were significantly higher than those in groups A and C at 4 and 8 weeks after operation ( P<0.05). Conclusion: Axial stress stimulation can promote the expression of SDF-1α in bone defect tissue, activate and regulate the CXCR4 signal collected by marrow mesenchymal stem cells, and accelerate bone regeneration in bone defect area.


Assuntos
Regeneração Óssea , Quimiocina CXCL12 , Cisteína , Animais , Fenômenos Biomecânicos , Quimiocina CXCL12/metabolismo , Cisteína/metabolismo , Masculino , Coelhos , Receptores CXCR4/metabolismo , Transdução de Sinais , Fator A de Crescimento do Endotélio Vascular/metabolismo
5.
Nat Commun ; 10(1): 1518, 2019 04 03.
Artigo em Inglês | MEDLINE | ID: mdl-30944331

RESUMO

When migrating in vivo, cells are exposed to numerous conflicting signals: chemokines, repellents, extracellular matrix, growth factors. The roles of several of these molecules have been studied individually in vitro or in vivo, but we have yet to understand how cells integrate them. To start addressing this question, we used the cephalic neural crest as a model system and looked at the roles of its best examples of positive and negative signals: stromal-cell derived factor 1 (Sdf1/Cxcl12) and class3-Semaphorins. Here we show that Sdf1 and Sema3A antagonistically control cell-matrix adhesion via opposite effects on Rac1 activity at the single cell level. Directional migration at the population level emerges as a result of global Semaphorin-dependent confinement and broad activation of adhesion by Sdf1 in the context of a biased Fibronectin distribution. These results indicate that uneven in vivo topology renders the need for precise distribution of secreted signals mostly dispensable.


Assuntos
Movimento Celular/fisiologia , Junções Célula-Matriz/fisiologia , Crista Neural/citologia , Animais , Adesão Celular/efeitos dos fármacos , Adesão Celular/fisiologia , Comunicação Celular/fisiologia , Linhagem Celular , Forma Celular/efeitos dos fármacos , Extensões da Superfície Celular/efeitos dos fármacos , Junções Célula-Matriz/efeitos dos fármacos , Junções Célula-Matriz/metabolismo , Quimiocina CXCL12/metabolismo , Feminino , Fibronectinas/metabolismo , Masculino , Manganês/metabolismo , Camundongos , Proteínas do Tecido Nervoso/fisiologia , Crista Neural/efeitos dos fármacos , Crista Neural/metabolismo , Receptores CXCR4/metabolismo , Semaforinas/metabolismo , Xenopus laevis/embriologia , Proteínas rac1 de Ligação ao GTP/metabolismo
6.
Nucleic Acids Res ; 47(11): 5465-5479, 2019 06 20.
Artigo em Inglês | MEDLINE | ID: mdl-31034558

RESUMO

Phosphorothioate-modified antisense oligonucleotides (PS-ASOs) interact with a host of plasma, cell-surface and intracellular proteins which govern their therapeutic properties. Given the importance of PS backbone for interaction with proteins, we systematically replaced anionic PS-linkages in toxic ASOs with charge-neutral alkylphosphonate linkages. Site-specific incorporation of alkyl phosphonates altered the RNaseH1 cleavage patterns but overall rates of cleavage and activity versus the on-target gene in cells and in mice were only minimally affected. However, replacing even one PS-linkage at position 2 or 3 from the 5'-side of the DNA-gap with alkylphosphonates reduced or eliminated toxicity of several hepatotoxic gapmer ASOs. The reduction in toxicity was accompanied by the absence of nucleolar mislocalization of paraspeckle protein P54nrb, ablation of P21 mRNA elevation and caspase activation in cells, and hepatotoxicity in mice. The generality of these observations was further demonstrated for several ASOs versus multiple gene targets. Our results add to the types of structural modifications that can be used in the gap-region to enhance ASO safety and provide insights into understanding the biochemistry of PS ASO protein interactions.


Assuntos
Membrana Celular/metabolismo , Citoplasma/metabolismo , Oligonucleotídeos Antissenso/química , Organofosfonatos/química , Oligonucleotídeos Fosforotioatos/química , Células 3T3-L1 , Animais , Caspases/metabolismo , Linhagem Celular , Quimiocina CXCL12/genética , Quimiocina CXCL12/metabolismo , Células HeLa , Hepatócitos/metabolismo , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Associadas à Matriz Nuclear/genética , Proteínas Associadas à Matriz Nuclear/metabolismo , Fatores de Transcrição de Octâmero/genética , Fatores de Transcrição de Octâmero/metabolismo , Oligonucleotídeos Antissenso/administração & dosagem , Oligonucleotídeos Fosforotioatos/administração & dosagem , Ligação Proteica , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Ribonuclease H/genética , Ribonuclease H/metabolismo , Receptores Depuradores Classe B/genética , Receptores Depuradores Classe B/metabolismo
7.
Cancer Biomark ; 25(1): 115-126, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31006667

RESUMO

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignancies, and its global morbidity and mortality are increasing. Previous studies confirmed that miR-342 was involved in the development and progression of malignant tumors. However, the relationship between miR-342 and Wnt/ß-catenin signaling pathway in HCC remains unknown. MATERIALS AND METHODS: Cell viability was detected by MTT assay. Immunofluorescence staining was used to detect Brdu-positive cells and Western blot was used to detect the apoptotic proteins. Furthermore, linear correlation analysis was used to investigate the possible relationship between miR-342 and the downstream genes of Wnt/ß-catenin signaling pathway in the progression of HCC. RESULTS: Over-expression of miR-342 significantly reduced cell proliferation and obviously increased apoptosis in HCC, while silencing of miR-342 showed an opposite effect on HCC cell proliferation and apoptosis. In addition, we found that the CXCL12 was the target gene of miR-342. This study also demonstrated that miR-342 up-regulation suppressed Wnt/ß-catenin signaling pathway by inhibiting CXCL12 expression. CONCLUSION: Up-regulation of miR-342 inhibited cell proliferation and induced cell apoptosis in HCC by inhibiting Wnt/ß-catenin signaling pathway, suggesting that miR-342 might act as a promising tumor gene therapeutic target for HCC patients.


Assuntos
Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/patologia , MicroRNAs/metabolismo , Via de Sinalização Wnt , beta Catenina/metabolismo , Apoptose , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Linhagem Celular , Proliferação de Células , Sobrevivência Celular , Quimiocina CXCL12/genética , Quimiocina CXCL12/metabolismo , Regulação para Baixo , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Masculino , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , Pessoa de Meia-Idade
8.
Turk J Haematol ; 36(2): 97-105, 2019 05 03.
Artigo em Inglês | MEDLINE | ID: mdl-30859801

RESUMO

Objective: Far beyond hemostasis and thrombosis, significant evidence has indicated the critical role of platelets in atherosclerosis. SDF-1 is among the pro-inflammatory chemokines that are increased in platelets of patients with coronary artery disease (CAD). The goal of the current work is to identify the in vitro effect of platelets from either CAD patients or healthy volunteers on the induction of macrophages and foam cells. Materials and Methods: The expression of SDF-1 on platelet surfaces in CAD patients and healthy volunteers was investigated using flow cytometry. We also evaluated the CXCR4/CXCR7 expression on monocytes from buffy coats of healthy volunteers. The effect of platelets from CAD patients and healthy volunteers on differentiation of monocytes and foam cell formation was evaluated using Oil Red O (ORO) staining. Flow cytometry and real-time PCR were also employed to evaluate surface markers and mRNA expression of genes involved in this process after co-culture of platelets with monocytes. Results: Monocytes in co-culture with platelets acquired a spindleshape appearance and ORO-positive lipid droplets. In addition, platelets could induce CD163 expression, as an important marker of M2 macrophage, and upregulate the mRNA expression of the SRB, CD36, ACAT, LXR-α, and ABCA1 genes in monocytes. Notably, platelets of CAD patients with higher expression of SDF-1, increased the expression of genes encoding SRB and CD36 as compared to platelets of healthy volunteers. Conclusion: Our results indicate that platelets from CAD patients could provoke monocyte differentiation into macrophages with an M2 phenotype, which in turn may participate in an atheroprotective process.


Assuntos
Plaquetas/patologia , Diferenciação Celular , Técnicas de Cocultura , Doença da Artéria Coronariana/patologia , Macrófagos/patologia , Monócitos/patologia , Aterosclerose/genética , Aterosclerose/metabolismo , Quimiocina CXCL12/genética , Quimiocina CXCL12/metabolismo , Doença da Artéria Coronariana/genética , Doença da Artéria Coronariana/metabolismo , Humanos
9.
Gene ; 700: 38-46, 2019 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-30898705

RESUMO

Anti-inflammation is a key process to restore tissue integrity and function. CXCL12 is a homeostasis chemokine, which plays a coordinating role in organogenesis, tumorigenesis and regeneration. In the present study we found that the uterus of abortion mice showed different histo-morphological changes with the development of abortion. The expression of chemokine CXCL12 and its receptor CXCR4 in abortion uterus showed a time-dependent pattern. Compared with normal pregnancy, the expression of CXCL12 and CXCR4 did not change in the uterus of GD7 abortion mice, but increased significantly in the uterus of GD8 and GD10 abortion mice. However, the expression of IFN-γ increased significantly in the uterus of GD7 abortion mice, while there was no significant change detected in GD8 aborted mice uterus. Our further data show that the expression of CXCL12 is not regulated by IFN-γ in endometrial stromal cell culture system in vitro. The treatment of CXCL12 significantly inhibits the expression of IFN-γ in in vitro cultured stromal cells and splenic monocytes. This suggests that CXCL12 may play an anti-inflammatory role in the uterus of abortion mice to promote the process of endometrial restoration after abortion, rather than participate in the process of abortion as a response molecule of IFN-γ.


Assuntos
Aborto Induzido/veterinária , Aborto Animal/metabolismo , Quimiocina CXCL12/genética , Quimiocina CXCL12/metabolismo , Interferon gama/efeitos adversos , Regulação para Cima , Aborto Animal/induzido quimicamente , Aborto Animal/genética , Animais , Células Cultivadas , Feminino , Camundongos , Gravidez , Receptores CXCR4/genética , Receptores CXCR4/metabolismo , Células Estromais/citologia , Células Estromais/efeitos dos fármacos , Células Estromais/metabolismo , Fatores de Tempo , Útero/citologia , Útero/efeitos dos fármacos , Útero/metabolismo
10.
Radiat Res ; 191(6): 518-526, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30925138

RESUMO

Low-dose radiation (LDR) has been confirmed to mobilize bone marrow-derived endothelial progenitor cells (EPCs) and promote diabetic wound healing. But it is unclear whether LDR acts directly on EPCs and promotes their proliferation and migration. Given the key role of advanced glycosylation end products (AGE) in the pathogenesis of diabetes, we used AGE to induce EPC damage. We then investigated the effect of LDR on the proliferation and migration of AGE-treated EPCs and explored the underlying mechanisms. EPCs cultured in vitro were treated with different concentrations of AGE, and the cells were then exposed to different low doses and treated with a specific antagonist for CXCR4, AMD3100 (1 lmol/l). The proliferation and migration abilities of EPCs were detected using the CCK-8 and wound healing assays, respectively. The mRNA and protein expression of SDF-1 and CXCR4 in AGE-treated EPCs were measured using qPCR and Western blot analysis, respectively. The expressions of ERK and phosphorylated ERK (pERK) were detected using Western blot analysis. The results showed that 200 mg/l and 400 mg/l AGE had an inhibitory effect on the proliferation of EPCs, and this inhibitory effect was exerted in a dose- and time-dependent manner. AGE significantly reduced the migration ability of EPCs cultured in vitro. After the cells received either 50 or 75 mGy low-dose irradiation, the proliferation of EPCs and AGE-treated EPCs was clearly increased; in addition, LDR also enhanced cell migration ability, but this enhancement was counteracted by AMD3100. Results from qPCR and Western blot analysis showed that LDR increased the mRNA and protein expression of SDF-1/ CXCR4. LDR also upregulated pERK expression in EPCs and AGE-treated EPCs, but LDR-induced upregulation of pERK expression was inhibited by AMD3100. These findings indicate that LDR can directly activate the SDF-1/CXCR4 biological axis and downstream ERK signaling pathway, and promote the proliferation and migration abilities of EPCs by increasing the expression of SDF-1, CXCR4 and pERK in EPCs.


Assuntos
Movimento Celular/efeitos dos fármacos , Movimento Celular/efeitos da radiação , Células Endoteliais/citologia , Produtos Finais de Glicação Avançada/farmacologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/efeitos da radiação , Células-Tronco/citologia , Animais , Células da Medula Óssea/citologia , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Quimiocina CXCL12/metabolismo , Relação Dose-Resposta à Radiação , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Masculino , Fenótipo , Ratos , Ratos Wistar , Receptores CXCR4/metabolismo , Células-Tronco/efeitos dos fármacos , Células-Tronco/efeitos da radiação
11.
DNA Cell Biol ; 38(5): 457-467, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30864829

RESUMO

Degenerative disc disease (DDD) is the main cause of low back pain, and the ingrowth of new blood vessels is one of its pathological features. The stromal cell-derived factor 1 (SDF1)/CXCR7 signaling axis plays a role in these physiological and pathological activities. The aims of this study were to explore whether this signaling axis participates in the angiogenesis of degenerated intervertebral discs (IVDs) and to define its underlying mechanism. In this study, we cocultured human nucleus pulposus cells (NPCs) and vascular endothelial cells (VECs) and regulated the expression of SDF1/CXCR7 to investigate the effect of VEC angiogenesis by NPCs. The results revealed that angiogenesis was enhanced with increased SDF1 and that angiogenesis was weakened with the inhibition of CXCR7. We found that PI3K/AKT was involved in the downstream pathway in the coculture. VEC angiogenesis induction by NPCs was enhanced with an increase in pAKT or a decrease in PTEN. We conclude that the SDF1/CXCR7 signaling axis plays a role in the angiogenesis of degenerated IVD through the PI3K/AKT pathway.


Assuntos
Quimiocina CXCL12/metabolismo , Degeneração do Disco Intervertebral/patologia , Neovascularização Patológica/patologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptores CXCR/metabolismo , Idoso , Células Cultivadas , Quimiocina CXCL12/genética , Técnicas de Cocultura , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Feminino , Humanos , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/metabolismo , Masculino , Pessoa de Meia-Idade , Neovascularização Patológica/genética , Neovascularização Patológica/metabolismo , Núcleo Pulposo/metabolismo , Núcleo Pulposo/patologia , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/genética , Receptores CXCR/genética , Transdução de Sinais
12.
Mol Med Rep ; 19(5): 3696-3706, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30896799

RESUMO

Epithelial­mesenchymal transition (EMT) has been demonstrated to serve a crucial role in the progression of interstitial fibrosis, which is one of the principal pathological features of chronic allograft nephropathy (CAN). However, to the best of our knowledge, the mechanisms of EMT in CAN have not been investigated. In the present study, the effect of stromal cell­derived factor 1 (SDF­1) and the Wnt signaling pathway on the progression of EMT following kidney transplantation was investigated. The CAN model was established using Fisher 344 and Lewis rats, treated with low­dose cyclosporine with or without AMD3100. CAN was confirmed by the pathological alterations and chronic allograft damage index scoring, and EMT was confirmed by western blotting and reverse transcription­quantitative polymerase chain reaction. In the AMD3100 group, there were lower expression levels of α­SMA and higher expression levels of E­cadherin, which indicated that CAN and EMT were ameliorated by AMD3100. The kidney tissue was analyzed using an mRNA + long noncoding (lnc)RNA microarray. A total of 506 mRNAs and 404 lncRNAs were demonstrated to be significantly differentially expressed between the two groups, which revealed the involvement of SDF­1/CXC chemokine receptor 4 (CXCR4) and the Wnt pathway. SDF­1 was demonstrated to induce EMT in vitro through the upregulation of α­SMA, downregulation of E­cadherin and the wound healing assay, and in the rat renal tubular epithelial cells via the nuclear accumulation of ß­catenin, which were all inhibited by either AMD3100 or DKK­1. CXXC finger protein 5 (CXXC5), a negative regulator of the Wnt pathway, was downregulated following treatment with SDF­1, which was inhibited by AMD3100 but not by DKK­1. Thus, CXXC5 may be a regulator downstream of SDF­1/CXCR4 in EMT. In conclusion, SDF­1/CXCR4 induces EMT of renal tubular epithelial cells with the involvement of the Wnt pathway, which may be a novel mechanism and therapeutic target in kidney allograft fibrosis of rats.


Assuntos
Quimiocina CXCL12/genética , Transição Epitelial-Mesenquimal/genética , Nefropatias/etiologia , Receptores CXCR4/genética , Transplante Homólogo/efeitos adversos , Via de Sinalização Wnt , Aloenxertos , Animais , Quimiocina CXCL12/metabolismo , Modelos Animais de Doenças , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Perfilação da Expressão Gênica , Compostos Heterocíclicos/farmacologia , Imuno-Histoquímica , Nefropatias/metabolismo , Transplante de Rim/efeitos adversos , Masculino , RNA Longo não Codificante/genética , Ratos , Receptores CXCR4/metabolismo
14.
Oxid Med Cell Longev ; 2019: 8148510, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30800209

RESUMO

Microenvironment plays a vital role in tumor progression; we focused on elucidating the role of hepatic stellate cells (HSCs) in hepatocarcinoma (HCC) aggressiveness and investigated the potential protective effect of curcumin on HSC-driven hepatocarcinoma angiogenesis and invasion. Our data suggest that HSCs increase HCC reactive oxygen species (ROS) production to upregulate hypoxia-inducible factor-1α (HIF-1α) expression to promote angiogenesis, epithelial to mesenchymal transition (EMT) process and invasion. And HSCs could secrete soluble factors, such as interleukin-6 (IL-6), vascular endothelial cell growth factor (VEGF), and stromal-derived factor-1 (SDF-1) to facilitate HCC progression. Curcumin could significantly suppress the above HSC-induced effects in HCC and could abrogate ROS and HIF-1α expression in HCC. HIF-1α or connective tissue growth factor (CTGF) knockdown could abolish the aforementioned curcumin affection. Moreover, CTGF is a downstream gene of HIF-1α. In addition, nuclear factor E2-related factor 2 (Nrf2) and glutathione (GSH) are involved in curcumin protection of HCC. These data indicate that curcumin may induce ROS scavenging by upregulating Nrf2 and GSH, thus inhibiting HIF-1α stabilization to suppress CTGF expression to exhibit its protection on HCC. Curcumin has a promising therapeutic effect on HCC. CTGF is responsible for curcumin-induced protection in HCC.


Assuntos
Carcinoma Hepatocelular/tratamento farmacológico , Fator de Crescimento do Tecido Conjuntivo/genética , Curcumina/uso terapêutico , Regulação para Baixo , Células Estreladas do Fígado/patologia , Neoplasias Hepáticas/tratamento farmacológico , Neovascularização Patológica/tratamento farmacológico , Carcinoma Hepatocelular/patologia , Quimiocina CXCL12/metabolismo , Fator de Crescimento do Tecido Conjuntivo/metabolismo , Curcumina/farmacologia , Regulação para Baixo/efeitos dos fármacos , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Glutationa/metabolismo , Células Hep G2 , Células Estreladas do Fígado/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Interleucina-6/metabolismo , Neoplasias Hepáticas/patologia , Metaloproteinase 9 da Matriz/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Invasividade Neoplásica , Neovascularização Patológica/patologia , Fator A de Crescimento do Endotélio Vascular/metabolismo
15.
J Int Med Res ; 47(3): 1264-1278, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30727793

RESUMO

OBJECTIVE: This study aimed to examine the role of spherical silica nanoparticles (SiNPs) on human bronchial epithelial (BEAS-2B) cells through inflammation. METHODS: Human mononuclear (THP-1) cells and BEAS-2B cells were co-cultured in transwell chambers and treated with 800 mmol/L benzo[ a]pyrene-7, 8-dihydrodiol-9, 10-epoxide (BPDE) and 12.5 µg/mL SiNPs for 24 hours. For controls, cells were treated with BPDE alone. Subcutaneous tumorigenicity and epithelial-mesenchymal transition (EMT) of BEAS-2B cells were measured. The cells were blocked with a stromal cell-derived factor-1α (SDF-1α)-specific antibody. EMT was analyzed in cells treated with 800 mmol/L BPDE and 12.5 µg/mL SiNPs relative to matched control cells and xenografts in vivo. Serum SDF-1α levels were measured in 23 patients with lung adenocarcinoma in Xuanwei, in 25 with lung adenocarcinoma outside Xuanwei, and in 22 with benign pulmonary lesions in Xuanwei. RESULTS: SiNPs significantly promoted tumorigenesis and EMT, induced the release of SDF-1α, and activated AKT (ser473) in BEAS-2B cells. EMT and phosphorylated AKT (ser473) and glycogen synthase kinase-3ß levels were decreased when blocked by SDF-1α antibody in BEAS-2B cells. SDF-1α was mainly secreted by THP-1 cells. CONCLUSION: SiNPs combined with BPDE promote EMT of BEAS-2B cells via the AKT pathway by inducing release of SDF-1α from THP-1 cells.


Assuntos
Brônquios/patologia , Transformação Celular Neoplásica/patologia , Quimiocina CXCL12/metabolismo , Transição Epitelial-Mesenquimal , Neoplasias Pulmonares/patologia , Nanopartículas/administração & dosagem , Dióxido de Silício/química , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Animais , Apoptose , Brônquios/metabolismo , Proliferação de Células , Transformação Celular Neoplásica/metabolismo , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Feminino , Humanos , Neoplasias Pulmonares/metabolismo , Masculino , Camundongos , Camundongos Nus , Pessoa de Meia-Idade , Nanopartículas/química , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de Xenoenxerto
16.
ACS Appl Mater Interfaces ; 11(9): 8878-8895, 2019 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-30777748

RESUMO

Electrospun scaffolds have been broadly studied to enhance bone regeneration because of the ability to simulate the structure and biological functions of the extracellular matrix. Polydopamine (PDA) is used to coat various surfaces at a slightly basic pH (8-8.5) and spontaneously reacts with nucleophilic functional groups. It is suitable for surface modifications of scaffolds correlated with bone formation. E7 is a newly discovered peptide with specific affinity for bone marrow mesenchymal stem cells (BMSCs). It can be useful for recruiting stem cells. Here, electrospun silk fibroin (SF) scaffolds were fabricated, and PDA was used for surface modification followed by grafting E7 (SF-PDA-E7). These composite SF-PDA-E7 electrospun scaffolds improved hydrophilicity, facilitated cell proliferation and adhesion, and boosted the osteogenic differentiation of BMSCs by creating osteoinduction conditions under the synergistic effects of PDA and E7. Moreover, the scaffolds showed high efficiency for recruiting BMSCs induced by E7 both in vitro and in vivo, which was associated with the SDF-1α/CXCR4 axis and the p38, extracellular signal-related kinase, and Akt signal transduction pathways. These functionalized electrospun scaffolds promoted regeneration of bone in the rat calvarial bone defect model. In general, this study verified that PDA could be a simple and efficient method for surface modification, and E7-grafted PDA-modified SF electrospun scaffolds were suitable for bone tissue engineering.


Assuntos
Regeneração Óssea , Fibroínas/química , Indóis/química , Peptídeos/química , Polímeros/química , Tecidos Suporte/química , Animais , Doenças Ósseas/patologia , Doenças Ósseas/terapia , Células da Medula Óssea/citologia , Osso e Ossos/diagnóstico por imagem , Osso e Ossos/patologia , Técnicas de Cultura de Células/instrumentação , Técnicas de Cultura de Células/métodos , Diferenciação Celular , Quimiocina CXCL12/metabolismo , Interações Hidrofóbicas e Hidrofílicas , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/patologia , Osteocalcina/genética , Osteocalcina/metabolismo , Osteogênese , Peptídeos/metabolismo , Ratos , Propriedades de Superfície , Engenharia Tecidual
17.
J Biol Chem ; 294(16): 6494-6505, 2019 04 19.
Artigo em Inglês | MEDLINE | ID: mdl-30808710

RESUMO

Missing in metastasis (MIM), an inverse Bin-Amphiphysin-Rvs (I-BAR) domain protein, promotes endocytosis of C-X-C chemokine receptor 4 (CXCR4) in mammalian cells. In response to the CXCR4 ligand stromal cell-derived factor 1 (SDF-1 or CXCL12), MIM associates with RAS-related GTP-binding protein 7 (RAB7) 30 min after stimulation. However, RAB7's role in MIM function remains undefined. Here we show that RNAi-mediated suppression of RAB7 expression in human HeLa cells has little effect on the binding of MIM to RAB5 and on the recruitment of CXCR4 to early endosomes but effectively abolishes MIM-mediated CXCR4 degradation, chemotactic response, and sorting into late endosomes and lysosomes. To determine whether I-BAR domain proteins interact with RAB7, we examined cells expressing insulin receptor tyrosine kinase substrate (IRTKS), an I-BAR domain protein bearing an Src homology 3 (SH3) domain. We observed that both MIM and IRTKS interact with RAB5 at an early response to SDF-1 and that IRTKS binds poorly to RAB7 but strongly to RAB11 at a later time point. Moreover, IRTKS overexpression reduced CXCR4 internalization and enhanced the chemotactic response to SDF-1. Interestingly, deletion of the SH3 domain in IRTKS abolished the IRTKS-RAB11 interaction and promoted CXCR4 degradation. Furthermore, the SH3 domain was required for selective targeting of MIM-IRTKS fusion proteins by both RAB7 and RAB11. Hence, to the best of our knowledge, our results provide first evidence that the SH3 domain is critical in the regulation of specific endocytic pathways by I-BAR domain proteins.


Assuntos
Endocitose , Proteínas dos Microfilamentos/metabolismo , Proteínas de Neoplasias/metabolismo , Proteólise , Receptores CXCR4/metabolismo , Proteínas rab de Ligação ao GTP/metabolismo , Quimiocina CXCL12/genética , Quimiocina CXCL12/metabolismo , Endossomos/genética , Endossomos/metabolismo , Células HeLa , Humanos , Lisossomos/genética , Lisossomos/metabolismo , Proteínas dos Microfilamentos/genética , Proteínas de Neoplasias/genética , Receptores CXCR4/genética , Proteínas rab de Ligação ao GTP/genética , Domínios de Homologia de src
18.
Mediators Inflamm ; 2019: 3231696, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30733641

RESUMO

The bone marrow (BM) is not only a reservoir of hematopoietic stem cells but a repository of immunological memory cells. Further characterizing BM-resident memory T cells would be helpful to reveal the complicated relationship between the BM and immunological memory. In this study, we identified CD122high stem cell antigen-1 (Sca-1) high B cell lymphoma 2 (Bcl-2) high CD4+ stem cell-like memory T cells (TSCMs) as a distinct memory T cell subset, which preferentially reside in the BM, where they respond vigorously to blood-borne antigens. Interestingly, the natural CD4+ TSCMs homing to the BM colocalized with VCAM-1+ IL-15+ IL-7+ CXCL-12+ stromal cells. Furthermore, compared to spleen-resident CD4+ TSCMs, BM-resident TSCMs induced the production of high-affinity antibodies against influenza by B lymphocytes more efficiently. Taken together, these observations indicate that the BM provides an appropriate microenvironment for the survival of CD4+ TSCMs, which broadens our knowledge regarding the memory maintenance of antigen-specific CD4+ T lymphocytes.


Assuntos
Anticorpos Antivirais/imunologia , Células da Medula Óssea/citologia , Linfócitos T CD4-Positivos/citologia , Memória Imunológica , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/terapia , Animais , Ataxina-1/metabolismo , Linfócitos T CD8-Positivos/citologia , Quimiocina CXCL12/metabolismo , Células-Tronco Hematopoéticas/citologia , Interleucina-15/metabolismo , Subunidade beta de Receptor de Interleucina-2/metabolismo , Interleucina-7/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Células Estromais , Molécula 1 de Adesão de Célula Vascular/metabolismo
19.
Thromb Haemost ; 119(2): 274-284, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30609443

RESUMO

Endothelial progenitor cells (EPCs) have been suggested to contribute to the neovascularization of infantile haemangioma (IH). There is strong evidence of the efficacy of propranolol in the treatment of IH, possibly by inhibiting both vasculogenesis and angiogenesis in the tumour. We evaluate the frequency of circulating endothelial colony forming cells (ECFCs), as the best EPC surrogate, in patients with IH at diagnosis and while receiving propranolol by an ex vivo 12-month longitudinal study. Biological aspects of the ECFCs, such as their in vitro angiogenic potential, membrane CXCR4 expression and Ca2+ signalling, were investigated. Circulating ECFCs were isolated by in vitro culture and expanded for 2 to 3 passages in 23 patients with IH (median age: 5.5 months, range: 5.5 weeks-11 months) before and 3, 6, 9 and 12 months after receiving propranolol. Twenty-four healthy subjects comparable for age were also assessed (CTRLs). Untreated patients with IH had a circulating ECFC frequency lower (p = 0.001) than CTRLs; nevertheless, in in vitro starving conditions, ECFCs showed enhanced capacity to form tube-like structures than those of CTRLs. Patients with IH following the therapy with propranolol had a significantly increased (p = 0.022) circulating ECFC frequency, that showed a diminished tube-like formation capacity in vitro, and an altered constitutive store-operated Ca2+ entry. ECFCs play a role in IH pathogenesis; the response to propranolol therapy is associated with their increased frequency in the peripheral blood and a reduction of their vasculogenic activity.


Assuntos
Células Endoteliais/citologia , Hemangioma/tratamento farmacológico , Hemangioma/metabolismo , Neovascularização Patológica , Propranolol/uso terapêutico , Antagonistas Adrenérgicos beta/uso terapêutico , Antígenos CD34/metabolismo , Cálcio/química , Sinalização do Cálcio , Movimento Celular , Quimiocina CXCL12/metabolismo , Células Endoteliais/efeitos dos fármacos , Células Progenitoras Endoteliais/citologia , Feminino , Citometria de Fluxo , Humanos , Lactente , Recém-Nascido , Cinética , Antígenos Comuns de Leucócito/metabolismo , Estudos Longitudinais , Masculino , Células-Tronco Mesenquimais/citologia , Neovascularização Fisiológica , Fenótipo , Receptores CXCR4/metabolismo
20.
Breast Cancer Res Treat ; 174(3): 679-691, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30632021

RESUMO

PURPOSE: Plasmacytoid dendritic cells (PDCs) infiltration into breast cancer tissues is associated with poor prognosis. Also, CXCR4 shows compelling evidences to be exploited by cancer cells to migrate to distant sites. The present study investigated lymph node metastasis in the light of PDCs infiltration and the potential cross talk with CXCR4/SDF-1 chemokine axis. METHODS: We assessed circulating PDCs proportions drained from the axillary tributaries, and the in situ expression of both CD303 and CXCR4 in breast cancer patients with positive lymph nodes (pLN) and negative lymph nodes (nLN) using immunohistochemistry and flow cytometry. We also analyzed the expression of SDF-1 in lymph nodes of pLN and nLN patients. We studied the effect of the secretome of PDCs of pLN and nLN patients on the expression of CXCR4 and activation of NF-κB in human breast cancer cell lines SKBR3 and MCF-7. TNF-α mRNA expression level in PDCs from both groups was determined by qPCR. RESULTS: Our findings indicate increased infiltration of PDCs in breast cancer tissues of pLN patients than nLN patients, which correlates with CXCR4+ cells percentage. Interestingly, SDF-1 is highly immunostained in lymph nodes of pLN patients compared to nLN patients. Our in vitro experiments demonstrate an upregulation of NF-κB expression and CXCR4 cells upon stimulation with PDCs secretome of pLN patients than those of nLN patients. Also, PDCs isolated from pLN patients exhibited a higher TNF-α mRNA expression than nLN patients. Treatment of MCF-7 cell lines with TNF-α significantly upregulates CXCR4 expression. CONCLUSIONS: Our findings suggest a potential role for microenvironmental PDCs in breast cancer lymph node metastasis via CXCR4/SDF-1 axis.


Assuntos
Neoplasias da Mama/patologia , Quimiocina CXCL12/metabolismo , Células Dendríticas/patologia , Metástase Linfática/patologia , NF-kappa B/metabolismo , Receptores CXCR4/metabolismo , Fator de Necrose Tumoral alfa/genética , Adulto , Idoso , Neoplasias da Mama/sangue , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Células Dendríticas/metabolismo , Feminino , Humanos , Metástase Linfática/genética , Células MCF-7 , Pessoa de Meia-Idade , Transdução de Sinais , Microambiente Tumoral
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