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1.
Medicine (Baltimore) ; 99(1): e18596, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31895808

RESUMO

Diabetic kidney disease (DKD) is a leading cause of end-stage renal disease. Because the molecular mechanisms of DKD are not fully understood, exploration of hub genes and the mechanisms underlying this disease are essential for elucidating the pathogenesis and progression of DKD. Accordingly, in this study, we performed an analysis of gene expression in DKD. The differentially expressed genes (DEGs) included 39 upregulated genes and 113 downregulated genes in the GSE30528 dataset and 127 upregulated genes and 18 downregulated genes in the GSE30529 dataset. Additionally, functional analyses were performed to determine the roles of DEGs using glomeruli samples from patients with DKD and healthy controls from the GSE30528 dataset and using tubule samples from patients with DKD and healthy controls from the GSE30529 dataset. These DEGs were enriched in pathways such as the Wnt signaling pathway, metabolic pathways, and the mammalian target of rapamycin signaling pathway in the GSE30528 dataset and the longevity regulating pathway and Ras signaling pathway in the GSE30529 dataset. Moreover, a protein-protein interaction network was constructed using the identified DEGs, and hub gene analysis was performed. Furthermore, correlation analyses between key genes and pathological characteristics of DKD indicated that CCR4, NTNG1, HGF and ISL1 are related to DKD, and NTNG1 and HGF may server as diagnostic biomarkers in DKD using the receiver-operator characteristic (ROC) curve. Collectively, our findings established 2 reliable biomarkers for DKD.


Assuntos
Nefropatias Diabéticas/metabolismo , Fator de Crescimento de Hepatócito/metabolismo , Rim/metabolismo , Netrinas/metabolismo , Biomarcadores/metabolismo , Estudos de Casos e Controles , Proteínas Ligadas por GPI/metabolismo , Humanos
2.
DNA Cell Biol ; 39(3): 451-458, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31910350

RESUMO

Gene delivery from tissue engineering scaffold is a novel strategy in regulating long-term growth and function of cells in vitro culture. In this study, a hepatocyte growth factor plasmid/polyetherimide (pHGF/PEI) polyplex delivering alginate (AL)/galactosylated chitosan (GC) (pHGF/PEI-AL/GC) sponge scaffold was prepared for the in vitro coculture of hepatocytes/3T3 cells. The pHGF/PEI polyplex released for 6 days in the sponge scaffold with weight ratio of AL/GC being 3:1 and fixed amount of pHGF being 40 µg (24-well scaffold). In addition, the 3T3 cells culturing in the pHGF/PEI-AL/GC sponge scaffold could be continually transfected and expressed the exogenous HGF for 6 days. Furthermore, the albumin secretion and urea synthesis of hepatocytes were significantly enhanced when cocultured with 3T3 cells in the pHGF/PEI-AL/GC sponge scaffold compared with that in the AL/GC sponge without pHGF. In summary, the preparation of AL/GC sponge scaffold delivering pHGF/PEI polyplex is a critical significance for maintaining the long-term survival and function of primary hepatocytes in vitro.


Assuntos
Quitosana/análogos & derivados , Técnicas de Transferência de Genes , Fator de Crescimento de Hepatócito/metabolismo , Hepatócitos/metabolismo , Tecidos Suporte/química , Células 3T3 , Alginatos/química , Animais , Células Cultivadas , Técnicas de Cocultura/métodos , Galactose/análogos & derivados , Fator de Crescimento de Hepatócito/genética , Hepatócitos/citologia , Hepatócitos/efeitos dos fármacos , Masculino , Camundongos , Polímeros/química , Ratos , Ratos Wistar , Tecidos Suporte/efeitos adversos
3.
Braz J Med Biol Res ; 53(1): e9144, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31939600

RESUMO

Wound scarring remains a major challenge for plastic surgeons. Transforming growth factor (TGF)-ß plays a key role in the process of scar formation. Previous studies have demonstrated that truncated TGF-ß type II receptor (t-TGF-ßRII) is unable to continue signal transduction but is still capable of binding to TGF-ß, thereby blocking the TGF-ß signaling pathway. Hepatocyte growth factor (HGF) is a multifunctional growth factor that promotes tissue regeneration and wound healing. Theoretically, the combination of HGF and t-TGF-ßRII would be expected to exert a synergistic effect on promoting wound healing and reducing collagen formation. In the present study, lentivirus-mediated transfection of the two genes (t-TGF-ßRII/HGF) into fibroblasts in vitro and in a rat model in vivo was used. The results demonstrated that the expression of t-TGF-ßRII and HGF in NIH-3T3 cells was successfully induced. The expression of both molecules significantly reduced collagen I and III expression, and also inhibited fibroblast proliferation. Furthermore, histological examination and scar quantification revealed less scarring in the experimental wound in a rat model. Moreover, on macroscopic inspection, the experimental wound exhibited less visible scarring compared with the control. Therefore, the present study demonstrated that the combination gene therapy of t-TGF-ßRII and HGF promoted wound healing, with less scarring and more epithelial tissue formation, not only by suppressing the overgrowth of collagen due to its antifibrotic effect, but also by promoting tissue regeneration.


Assuntos
Cicatriz/metabolismo , Colágeno/metabolismo , Fator de Crescimento de Hepatócito/metabolismo , Transfecção , Fator de Crescimento Transformador beta2/metabolismo , Animais , Proliferação de Células , Cicatriz/patologia , Camundongos , Modelos Animais , Ratos , Ratos Sprague-Dawley
4.
Cancer Sci ; 111(2): 406-417, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31785057

RESUMO

STMN1 has been regarded as an oncogene and its upregulation is closely associated with malignant behavior and poor prognosis in multiple cancers. However, the detailed functions and underlying mechanisms of STMN1 are still largely unknown in hepatocellular carcinoma (HCC) development. Herein, we analyzed STMN1 expression and the related clinical significance in HCC by using well-established Protein Atlas, The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) cancer databases. Analysis indicated that STMN1 was highly expressed in HCC and closely associated with vascular invasion, higher histological grade, advanced clinical grade and shorter survival time in HCC patients. Overexpressing and silencing STMN1 in HCC cell lines showed that STMN1 could regulate cell proliferation, migration, drug resistance, cancer stem cell properties in vitro as well as tumor growth in vivo. Further experiments showed that STMN1 mediated intricate crosstalk between HCC and hepatic stellate cells (HSC) by triggering the hepatocyte growth factor (HGF)/MET signal pathway. When HSC were cocultured with HCC cells, HSC secreted more HGF to stimulate the expression of STMN1 in HCC cells. Mutually, STMN1 upregulation in HCC cells facilitated HSC activation to acquire cancer-associated fibroblast (CAF) features. The MET inhibitor crizotinib significantly blocked this crosstalk and slowed tumor growth in vivo. In conclusion, our findings shed new insight on STMN1 function, and suggest that STMN1 may be used as a potential marker to identify patients who may benefit from MET inhibitor treatment.


Assuntos
Carcinoma Hepatocelular/patologia , Células Estreladas do Fígado/citologia , Neoplasias Hepáticas/patologia , Transdução de Sinais , Estatmina/genética , Animais , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Linhagem Celular Tumoral , Técnicas de Cocultura , Resistencia a Medicamentos Antineoplásicos , Retroalimentação Fisiológica , Regulação Neoplásica da Expressão Gênica , Células Estreladas do Fígado/metabolismo , Fator de Crescimento de Hepatócito/metabolismo , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Masculino , Camundongos , Gradação de Tumores , Transplante de Neoplasias , Células-Tronco Neoplásicas/metabolismo , Proteínas Proto-Oncogênicas c-met/metabolismo , Regulação para Cima
5.
Anticancer Res ; 39(12): 6685-6691, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31810933

RESUMO

BACKGROUND/AIM: No effective therapeutics have yet been developed for pancreatic cancer. 2-Methoxy-4-vinyl phenol (2M4VP), a member of the class of phenols, has been demonstrated to have anti-inflammatory properties and cause cell cycle arrest making it an attractive candidate drug for the treatment of pancreatic cancer. MATERIALS AND METHODS: The effects of 2M4VP were examined in Panc-1 and SNU-213 human pancreatic cancer cells. RESULTS: 2M4VP had anticancer effects on pancreatic cancer cell lines, Panc-1 and SNU-213. 2M4VP reduced the viability of Panc-1 cells by inhibiting the expression of the cell nuclear antigen (PCNA) protein. 2M4VP also suppressed the migratory activity of both cell lines. In addition, treatment with 2M4VP effectively decreased the phosphorylation of Focal Adhesion Kinase (FAK) and AKT. CONCLUSION: 2M4VP might be used as a pancreatic cancer treatment supplement.


Assuntos
Movimento Celular/efeitos dos fármacos , Quinase 1 de Adesão Focal/antagonistas & inibidores , Guaiacol/análogos & derivados , Neoplasias Pancreáticas/tratamento farmacológico , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Compostos de Vinila/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Quinase 1 de Adesão Focal/metabolismo , Guaiacol/farmacologia , Fator de Crescimento de Hepatócito/metabolismo , Humanos , Proteínas de Neoplasias/antagonistas & inibidores , Proteínas de Neoplasias/metabolismo , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/patologia , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos
6.
Elife ; 82019 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-31535973

RESUMO

In most vertebrates, the upper digestive tract is composed of muscularized jaws linked to the esophagus that permits food ingestion and swallowing. Masticatory and esophagus striated muscles (ESM) share a common cardiopharyngeal mesoderm (CPM) origin, however ESM are unusual among striated muscles as they are established in the absence of a primary skeletal muscle scaffold. Using mouse chimeras, we show that the transcription factors Tbx1 and Isl1 are required cell-autonomously for myogenic specification of ESM progenitors. Further, genetic loss-of-function and pharmacological studies point to MET/HGF signaling for antero-posterior migration of esophagus muscle progenitors, where Hgf ligand is expressed in adjacent smooth muscle cells. These observations highlight the functional relevance of a smooth and striated muscle progenitor dialogue for ESM patterning. Our findings establish a Tbx1-Isl1-Met genetic hierarchy that uniquely regulates esophagus myogenesis and identify distinct genetic signatures that can be used as framework to interpret pathologies arising within CPM derivatives.


Assuntos
Padronização Corporal , Esôfago/embriologia , Regulação da Expressão Gênica no Desenvolvimento , Mesoderma/embriologia , Músculo Estriado/embriologia , Animais , Fator de Crescimento de Hepatócito/metabolismo , Proteínas com Homeodomínio LIM/metabolismo , Camundongos , Proteínas Proto-Oncogênicas c-met/metabolismo , Transdução de Sinais , Proteínas com Domínio T/metabolismo , Fatores de Transcrição/metabolismo
7.
Radiat Res ; 192(5): 517-526, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31442107

RESUMO

At low doses, ionizing radiation activates endothelial cells and promotes angiogenesis. However, it is still unknown if other cells may contribute to this process. In this study, the effect of low-dose ionizing radiation (LDIR) in modulating the pro-angiogenic potential of adipocytes was investigated. Adipocytes are known to secrete multiple angiogenic factors and adipokines that induce angiogenesis. In this work, a confluent monolayer of 3T3-L1 pre-adipocytes was exposed to low doses (0.1 and 0.3 Gy) and to higher doses (0.5, 0.8 and 1.0 Gy), as control. Our data show that the adipocyte-conditioned media (A-CM) from mature adipocytes differentiated from low-dose irradiated pre-adipocytes presented a higher angiogenic potential, compared to mature adipocytes differentiated from sham-irradiated control preadipocytes. The vascular endothelial growth factor (VEGF)-A levels were significantly increased in A-CM from the 0.1 Gy (P < 0.05) and 0.3 Gy (P < 0.01) experimental conditions and a significant increase was found in response to 0.3 Gy dose of radiation for VEGF-C, angiopoietin-2 (ANG-2) and hepatocyte growth factor (HGF). Moreover, 0.3 Gy dose of radiation significantly increased the expression of matrix metalloproteinase (MMP)-2 active forms. In vitro, the A-CM from the 0.1 and 0.3 Gy doses experimental conditions significantly accelerated endothelial cell migration after an in vitro wound healing assay. Importantly, in vivo, the A-CM corresponding to the 0.3 Gy experimental condition significantly induced the growth of more blood vessels towards the inoculation area in the chick embryo chorioallantoic membrane (CAM). In conclusion, this work reveals a new mechanism by which low-dose radiation might promote angiogenesis, enhancing the angiogenic potential of A-CM.


Assuntos
Adipócitos/efeitos da radiação , Meios de Cultivo Condicionados/química , Neovascularização Fisiológica , Radiação Ionizante , Células 3T3-L1 , Adipócitos/metabolismo , Angiopoietina-1/metabolismo , Animais , Diferenciação Celular , Movimento Celular/efeitos da radiação , Células Cultivadas , Embrião de Galinha , Membrana Corioalantoide/metabolismo , Relação Dose-Resposta à Radiação , Fator de Crescimento de Hepatócito/metabolismo , Células Endoteliais da Veia Umbilical Humana/efeitos da radiação , Humanos , Camundongos , Fator A de Crescimento do Endotélio Vascular/metabolismo , Fator C de Crescimento do Endotélio Vascular/metabolismo
8.
Int J Med Sci ; 16(6): 854-863, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31337959

RESUMO

Background: HGF/MET pathway may have a role in pulmonary hypertension (PH). However, the link between the pathway and development of target organ damage in PH remains elusive. We aimed to demonstrate the relation between plasma HGF and HGF/MET tissue expressions in affected organs during PH progression. Methods: 12 weeks old male Wistar rats were injected with monocrotaline (MCT, 60 mg/kg, s.c.) to induce PH and sacrificed after 1, 2 and 4 weeks. Controls received saline. mRNA levels of HGF regulatory complex (Hgf, Met, Hgfa, Hai-1, Hai-2) were determined in right and left ventricles (RV, LV), lungs, pulmonary artery and liver by RT-qPCR. HGF protein levels in plasma were analysed by ELISA. Results: PH development was associated with a progressive elevation of HGF plasma levels that correlated with relative RV mass. Furthermore, Hgf mRNA expressions at week 4 were upregulated solely in the cardiac ventricles while being downregulated in a. pulmonalis, lungs and liver. Met and Hai-1/Hai-2 followed a similar pattern and were upregulated in cardiac ventricles, where Hgfa remained unchanged, but downregulated in lungs. Conclusion: We suggest that cardiac overexpression of Hgf might contribute to increased plasma HGF in MCT-induced PH. HGF could be exploited as a cardiospecific biomarker and HGF/MET pathway as a target in drug discovery for PH.


Assuntos
Insuficiência Cardíaca/diagnóstico , Ventrículos do Coração/patologia , Fator de Crescimento de Hepatócito/metabolismo , Hipertensão Pulmonar/complicações , Remodelação Ventricular , Animais , Biomarcadores/sangue , Biomarcadores/metabolismo , Modelos Animais de Doenças , Progressão da Doença , Regulação para Baixo , Insuficiência Cardíaca/sangue , Insuficiência Cardíaca/etiologia , Fator de Crescimento de Hepatócito/sangue , Humanos , Hipertensão Pulmonar/sangue , Hipertensão Pulmonar/induzido quimicamente , Masculino , Monocrotalina/toxicidade , Proteínas Proto-Oncogênicas c-met/metabolismo , Ratos , Ratos Wistar , Regulação para Cima
9.
Environ Toxicol ; 34(11): 1236-1245, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31313457

RESUMO

Hepatocyte growth factor (HGF) has recently been reported to exhibit antioxidant and antiapoptotic effects. Therefore, we investigated the effect of overexpression of HGF gene in H2 O2 -treated mesenchymal stem cells (MSCs). HGF-overexpression increased the cell viability from 50% to 84%, decreased the population of apoptotic cells from 20% to 16%, and decreased the intracellular reactive oxygen species (ROS) levels from 127% to 100% in cells treated with H2 O2 . HGF suppression decreased the cell viability from 58% to 36%, increased the population of apoptotic cells from 23 to 81%, and increased the intracellular ROS levels from 181% to 240% in cells exposed to H2 O2 . HGF-overexpression also reduced the expression levels of proapoptotic proteins in MSCs treated with H2 O2 . Phosphorylation of extracellular signal-regulated kinases, c-Jun amino-terminal kinases, and p38, which was induced by H2 O2 , decreased in MSCs overexpressing the HGF gene. Taken together, our results suggest that HGF has a protective effect on H2 O2 -induced apoptosis in MSCs.


Assuntos
Apoptose/efeitos dos fármacos , Fator de Crescimento de Hepatócito/metabolismo , Peróxido de Hidrogênio/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Sangue Fetal/citologia , Fator de Crescimento de Hepatócito/genética , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Espécies Reativas de Oxigênio/metabolismo
10.
Bull Exp Biol Med ; 167(3): 413-417, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31350657

RESUMO

A stimulating effect of a combination of hepatocyte growth factor (HGF) and glial neurotrophic factor (GDNF) on the growth of neurites in the spinal ganglion model was demonstrated. The mechanism of neurite growth in the spinal ganglion model is associated with transactivation of HGF c-met receptor in the presence of both HGF and GDNF. The combination of HGF and GDNF significantly activated mitogenic signaling cascade mediated by protein kinases ERK1/2, which can be a mechanism for increasing the number of neurites. Our findings can be used for developing effective methods for restoring impaired peripheral nerve function after traumatic and ischemic injury using a combination of GDNF and HGF.


Assuntos
Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Fator de Crescimento de Hepatócito/metabolismo , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Neuritos/metabolismo , Animais , Linhagem Celular Tumoral , Sistema de Sinalização das MAP Quinases/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Regeneração Nervosa/fisiologia , Fosforilação , Proteínas Proto-Oncogênicas c-met/metabolismo
11.
Cancer Sci ; 110(10): 3340-3349, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31342590

RESUMO

Aberrant activation of the MET/hepatocyte growth factor (HGF) receptor participates in the malignant behavior of cancer cells, such as invasion-metastasis and resistance to molecular targeted drugs. Many mutations in the MET extracellular region have been reported, but their significance is largely unknown. Here, we report the dysregulation of mutant MET originally found in a lung cancer patient with Val370 to Asp370 (V370D) replacement located in the extracellular SEMA domain. MET-knockout cells were prepared and reconstituted with WT-MET or V370D-MET. HGF stimulation induced MET dimerization and biological responses in cells reconstituted with WT-MET, but HGF did not induce MET dimerization and failed to induce biological responses in V370D-MET cells. The V370D mutation abrogated HGF-dependent drug resistance of lung cancer cells to epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKI). Compared with WT-MET cells, V370D-MET cells showed different activation patterns in receptor tyrosine kinases upon exposure to survival/growth-stressed conditions. Surface plasmon resonance analysis indicated that affinity between the extracellular region of V370D-MET and HGF was reduced compared with that for WT-MET. Further analysis of the association between V370D-MET and the separate domains of HGF indicated that the SP domain of HGF was unchanged, but its association with the NK4 domain of HGF was mostly lost in V370D-MET. These results indicate that the V370D mutation in the MET receptor impairs the functional association with HGF and is therefore a loss-of-function mutation. This mutation may change the dependence of cancer cell growth/survival on signaling molecules, which may promote cancer cell characteristics under certain conditions.


Assuntos
Fator de Crescimento de Hepatócito/metabolismo , Neoplasias Pulmonares/genética , Mutação de Sentido Incorreto , Proteínas Proto-Oncogênicas c-met/química , Proteínas Proto-Oncogênicas c-met/genética , Animais , Células CHO , Linhagem Celular Tumoral , Cricetulus , Resistencia a Medicamentos Antineoplásicos , Técnicas de Inativação de Genes , Humanos , Mutação com Perda de Função , Domínios Proteicos , Inibidores de Proteínas Quinases/farmacologia , Multimerização Proteica , Proteínas Proto-Oncogênicas c-met/metabolismo , Ativação Transcricional
12.
J Mol Histol ; 50(5): 405-415, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31256303

RESUMO

Viral myocarditis has been found to be one of the leading causes of sudden death in young adults. However, no effective drugs have been developed to intervene the progression of myocarditis. Accordingly, the present study is carried out to explore the protective role played by melatonin in the setting of viral myocarditis with a focus on Mst1-Hippo pathway, mitochondrial dysfunction and ER stress. Cardiac function was determined via echocardiographic examination. Mitochondrial function and ER stress were detected via ELISA, western blots, and immunofluorescence. Our data demonstrated that virus injection induced cardiac dysfunction as evidenced by reduced contractile function in myocardium. Besides, LDH release assay and western blotting analysis demonstrated that cardiomyocyte death was activated by virus injection. Interestingly, melatonin treatment improved cardiac function and repressed virus-mediated cardiomyocyte apoptosis. At the molecular levels, mitochondrial dysfunction was induced by virus infection, as indicated by mitochondrial membrane potential reduction, mPTP opening rate elevation and caspase-9-related apoptosis activation. Besides, ER stress parameters were also elevated in virus-treated cardiomyocytes. Interestingly, melatonin treatment maintained mitochondrial dysfunction and repressed ER stress. To the end, we found that Mst1 was upregulated by virus infection; this effect was attenuated through supplementation with melatonin. However, Mst1 overexpression reduced the beneficial impact exerted by melatonin on cardiomyocyte viability, mitochondrial function and ER homeostasis. Our study illustrated that melatonin treatment attenuated viral myocarditis via sustaining cardiomyocyte viability, repressing mitochondrial dysfunction and inhibiting ER stress in a manner dependent on Mst1 inhibition.


Assuntos
Estresse do Retículo Endoplasmático/efeitos dos fármacos , Fator de Crescimento de Hepatócito/antagonistas & inibidores , Melatonina/farmacologia , Mitocôndrias/efeitos dos fármacos , Miocardite/prevenção & controle , Proteínas Proto-Oncogênicas/antagonistas & inibidores , Animais , Sobrevivência Celular/efeitos dos fármacos , Fator de Crescimento de Hepatócito/metabolismo , Humanos , Mitocôndrias/patologia , Miócitos Cardíacos/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Proteínas Proto-Oncogênicas/metabolismo , Vírus/patogenicidade
13.
J Exp Clin Cancer Res ; 38(1): 270, 2019 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-31221203

RESUMO

Gliomas represent the most common type of malignant brain tumor, among which, glioblastoma remains a clinical challenge with limited treatment options and dismal prognosis. It has been shown that the dysregulated receptor tyrosine kinase (RTK, including EGFR, MET, PDGFRα, ect.) signaling pathways have pivotal roles in the progression of gliomas, especially glioblastoma. Increasing evidence suggests that expression levels of the RTK MET and its specific stimulatory factors are significantly increased in glioblastomas compared to those in normal brain tissues, whereas some negative regulators are found to be downregulated. Mutations in MET, as well as the dysregulation of other regulators of cross-talk with MET signaling pathways, have also been identified. MET and its ligand hepatocyte growth factor (HGF) play a critical role in the proliferation, survival, migration, invasion, angiogenesis, stem cell characteristics, and therapeutic resistance and recurrence of glioblastomas. Therefore, combined targeted therapy for this pathway and associated molecules could be a novel and attractive strategy for the treatment of human glioblastoma. In this review, we highlight progress made in the understanding of MET signaling in glioma and advances in therapies targeting HGF/MET molecules for glioma patients in recent years, in addition to studies on the expression and mutation status of MET.


Assuntos
Neoplasias Encefálicas/genética , Glioma/genética , Fator de Crescimento de Hepatócito/metabolismo , Proteínas Proto-Oncogênicas c-met/genética , Neoplasias Encefálicas/metabolismo , Movimento Celular , Proliferação de Células , Resistencia a Medicamentos Antineoplásicos , Regulação Neoplásica da Expressão Gênica , Glioma/metabolismo , Humanos , Mutação , Invasividade Neoplásica , Prognóstico , Proteínas Proto-Oncogênicas c-met/metabolismo , Transdução de Sinais , Regulação para Cima
14.
Growth Factors ; 37(1-2): 68-75, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-31185750

RESUMO

Biliary atresia (BA) is characterized by progressive destruction of the biliary system leading to liver fibrosis and deterioration of liver function. Serum hepatocyte growth factor (HGF) has been shown to be increased in cirrhotic diseases including BA. The aim of this study was to investigate the prognostic value of HGF levels in sera and liver tissue for the further disease course. A total of 49 serum and liver samples from infants with BA were acquired during Kasai-portoenterostomy (KPE) and analyzed by multiplex immunoassay including HGF, as marker of liver regeneration, and Interleukin 6 (IL-6) as a marker of inflammation. Both mediators showed no correlation with the outcome defined as favorable (survival with native liver (SNL)) or, in contrast, rapid deterioration of liver function requiring transplantation. Our data suggest that the degree of liver regeneration indicated by high levels of HGF within the liver is a dismissible factor in the post-KPE disease course.


Assuntos
Atresia Biliar/sangue , Fator de Crescimento de Hepatócito/sangue , Fígado/metabolismo , Portoenterostomia Hepática/efeitos adversos , Complicações Pós-Operatórias/sangue , Atresia Biliar/metabolismo , Atresia Biliar/cirurgia , Biomarcadores/sangue , Biomarcadores/metabolismo , Feminino , Fator de Crescimento de Hepatócito/metabolismo , Humanos , Lactente , Recém-Nascido , Interleucina-6/sangue , Interleucina-6/metabolismo , Masculino , Complicações Pós-Operatórias/metabolismo
15.
Acta Biomater ; 94: 330-339, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31176843

RESUMO

Human amniotic membrane (AM) has intrinsic anti-inflammatory, anti-fibrotic and antimicrobial properties. Tissue preservation methods have helped to overcome the short shelf life of fresh AM allowing "on demand" use of AM grafts. Cryopreserved AM that retains all native tissue components, including viable cells, has clinical benefits in treating chronic wounds. However, cryopreservation requires ultra-low temperature storage, limiting the use of cryopreserved products. To overcome this limitation, a new lyopreservation method has been developed for ambient storage of living tissues. The goal of this study was to investigate the viability and functionality of AM cells following lyopreservation. Fresh AM and devitalized lyopreserved AM (DLAM) served as positive and negative controls, respectively. Using live/dead staining, we confirmed the presence of living cells in viable lyopreserved AM (VLAM) and showed that these cells persisted up to 21 days in culture medium. The functionality of cells in VLAM was assessed by their differentiation potential and anti-fibrotic activity in vitro. With osteogenic induction, cells in VLAM deposited calcium within the membrane, a marker of osteogenic cells, in a time-dependent manner. The migration of human lung fibrotic fibroblasts in a scratch wound assay was reduced significantly in the presence of VLAM-derived conditioned medium. Quantitative PCR analyses indicated that VLAM reduced the expression of pro-fibrotic factors such as type I collagen and increased the expression of anti-fibrotic factors such as hepatocyte growth factor and anti-fibrotic microRNA in fibrotic fibroblasts. Taken together, these results demonstrate that endogenous cells in VLAM remain viable and functional post-lyophilization. STATEMENT OF SIGNIFICANCE: This study, for the first time, provides direct evidence showing that tissue viability and functional cells can be preserved by lyophilization. Similar to fresh amniotic membrane (AM), viable lyopreserved AM (VLAM) retains viable cells for extended periods of time. More importantly, these cells are functional and maintain their osteogenic differentiation potential and anti-fibrotic activity. Our results confirmed that the novel lyophilization method preserves tissue viability.


Assuntos
Âmnio/fisiologia , Fibroblastos/fisiologia , Fibrose/prevenção & controle , Liofilização/métodos , Diferenciação Celular , Linhagem Celular , Membrana Celular/metabolismo , Sobrevivência Celular , Criopreservação/métodos , Meios de Cultivo Condicionados/metabolismo , Fibrose/metabolismo , Regulação da Expressão Gênica , Fator de Crescimento de Hepatócito/metabolismo , Humanos , MicroRNAs/metabolismo , Osteogênese , Sobrevivência de Tecidos
16.
Int J Mol Sci ; 20(12)2019 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-31212972

RESUMO

Hepatocyte growth factor (HGF) is secreted as an inactive single-chain HGF (scHGF); however, only proteolytically processed two-chain HGF (tcHGF) can activate the MET receptor. We investigated the localization of tcHGF and activated/phosphorylated MET (pMET) using a tcHGF-specific antibody. In day 16.5 mouse embryos, total HGF (scHGF + tcHGF) was mainly localized in smooth muscle cells close to, but separate from, MET-positive epithelial cells in endodermal organs, including the stomach. In the adult stomach, total HGF was localized in smooth muscle cells, and tcHGF was mainly localized in the glandular base region. Immunostaining for pMET and Lgr5-driven green fluorescent protein (GFP) indicated that pMET localization overlapped with Lgr5+ gastric stem cells. HGF promoted organoid formation similar to EGF, indicating the potential for HGF to promote the survival and growth of gastric stem cells. pMET and tcHGF localizations changed during regeneration following gastric injury. These results indicate that MET is constantly activated in gastric stem cells and that the localization of pMET differs from the primary localization of precursor HGF but has a close relationship to tcHGF. Our results suggest the importance of the microenvironmental generation of tcHGF in the regulation of development, regeneration, and stem cell behavior.


Assuntos
Fator de Crescimento de Hepatócito/metabolismo , Organogênese , Cicatrização , Animais , Biomarcadores , Fator de Crescimento de Hepatócito/genética , Humanos , Imuno-Histoquímica , Camundongos , Camundongos Transgênicos , Miócitos de Músculo Liso/citologia , Miócitos de Músculo Liso/metabolismo , Organogênese/genética , Fosforilação , Transporte Proteico , Proteínas Proto-Oncogênicas c-met/metabolismo , Regeneração , Células-Tronco/citologia , Células-Tronco/metabolismo , Cicatrização/genética
17.
Biochem J ; 476(10): 1419-1431, 2019 05 21.
Artigo em Inglês | MEDLINE | ID: mdl-31036720

RESUMO

Phosphatase of regenerating liver (PRL) is overexpressed in metastatic cancers and actively drives their malignant progression. Many studies on cultured cancer cells have implied PRL overexpression as a stimulant for cellular signaling involved in cell proliferation. However, its role in the tightly adhered and polarized epithelial cells remains largely uncharacterized. In this study, we show that inducible expression of PRL in MDCK normal epithelial cells sensitized MET, the receptor for hepatocyte growth factor (HGF), to functional activation by HGF. We found that PRL expression amplified tyrosine phosphorylation levels of various proteins, among which MET was identified to be the most abundant. This phosphorylation occurred selectively at Y1234/1235 in the activation loop of MET, whereas phosphorylation of Y1349 in the effector-binding site, which is directly involved in downstream signaling, was almost undetectable. Consistently, PRL overexpression by itself did not cause observable alterations at the cellular level. However, when cells were stimulated with HGF, phosphorylation of Y1349 was much more strongly induced in PRL-expressing cells than in control cells. This resulted in robust cell scattering and tubulogenesis, even with low levels of HGF. Collectively, these results demonstrate a unique role of PRL in regulating MET function, which is known to be crucial for remodeling of epithelial tissues and malignant progression of cancers.


Assuntos
Fator de Crescimento de Hepatócito/metabolismo , Neoplasias/metabolismo , Proteínas Tirosina Fosfatases/metabolismo , Proteínas Proto-Oncogênicas c-met/metabolismo , Animais , Cães , Fator de Crescimento de Hepatócito/genética , Células Madin Darby de Rim Canino , Neoplasias/genética , Fosforilação , Estrutura Secundária de Proteína , Proteínas Tirosina Fosfatases/genética , Proteínas Proto-Oncogênicas c-met/genética
18.
Nat Chem Biol ; 15(6): 598-606, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31101918

RESUMO

Activation of hepatocyte growth factor (HGF) by proteolytic processing is triggered in cancer microenvironments, and subsequent signaling through the MET receptor is involved in cancer progression. However, the structure of HGF remains elusive, and few small/medium-sized molecules can modulate HGF. Here, we identified HiP-8, a macrocyclic peptide consisting of 12 amino acids, which selectively recognizes active HGF. Biochemical analysis and real-time single-molecule imaging by high-speed atomic force microscopy demonstrated that HiP-8 restricted the dynamic domains of HGF into static closed conformations, resulting in allosteric inhibition. Positron emission tomography using HiP-8 as a radiotracer enabled noninvasive visualization and simultaneous inhibition of HGF-MET activation status in tumors in a mouse model. Our results illustrate the conformational change in proteolytic activation of HGF and its detection and inhibition by a macrocyclic peptide, which may be useful for diagnosis and treatment of cancers.


Assuntos
Fator de Crescimento de Hepatócito/análise , Compostos Macrocíclicos/química , Neoplasias Experimentais/diagnóstico por imagem , Imagem Óptica , Peptídeos/química , Animais , Fator de Crescimento de Hepatócito/antagonistas & inibidores , Fator de Crescimento de Hepatócito/metabolismo , Compostos Macrocíclicos/farmacologia , Camundongos , Neoplasias Experimentais/tratamento farmacológico , Peptídeos/farmacologia , Tomografia por Emissão de Pósitrons
19.
Int J Mol Med ; 44(1): 227-239, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31115492

RESUMO

Endoplasmic reticulum (ER) stress in alveolar epithelial cells (AECs) is associated with the pathogenesis of pulmonary fibrosis. Bone marrow­derived mesenchymal stromal cells (BM­MSCs) can exert protective effects on ER­stressed AECs via paracrine signaling. In the present study, mouse lung epithelial (MLE)­12 cells were directly stimulated with various concentrations of bleomycin (BLM). MLE­12 cell apoptosis was detected by flow cytometry, and Ki67 expression was detected by immunofluorescence to reflect cell proliferation. The results revealed that BLM increased the protein expression levels of X­box binding protein 1 and immunoglobulin heavy chain­binding protein, thus inducing ER stress, and caused cell dysfunction by inhibiting proliferation and promoting apoptosis. In addition, MSC­derived conditioned medium (MSC­CM) protected MLE­12 cells from BLM­induced injury, by reducing ER stress, promoting cell proliferation and inhibiting cell apoptosis. Our previous studies reported that N­methyl­D­aspartate (NMDA) receptor activation partially inhibits the antifibrotic effect of BM­MSCs on BLM­induced pulmonary fibrosis through downregulating the paracrine factor hepatocyte growth factor (HGF). In the present study, the synthesis and secretion of HGF were detected by western blotting and ELISA, respectively. Results further demonstrated that NMDA inhibited the synthesis and secretion of HGF in BM­MSCs, and NMDA­preconditioned MSC­CM had no protective effects on BLM­induced injury in MLE­12 cells. In addition, activation of the NMDA receptor decreased the phosphorylation levels of extracellular signal­regulated kinase (ERK)1/2 in BM­MSCs. Using Honokiol and FR180204, the activator and inhibitor of ERK1/2, respectively, it was then revealed that Honokiol partially eliminated the decrease in HGF expression, whereas FR180204 further promoted the reduction in HGF caused by NMDA. Collectively, these findings suggested that NMDA receptor activation may downregulate HGF by inhibiting ERK signaling in BM­MSCs, thus weakening their protective effects on BLM­induced lung epithelial cell damage.


Assuntos
Células Epiteliais Alveolares/metabolismo , Bleomicina/efeitos adversos , Células da Medula Óssea/metabolismo , Fator de Crescimento de Hepatócito/metabolismo , Sistema de Sinalização das MAP Quinases , Células-Tronco Mesenquimais/metabolismo , Comunicação Parácrina , Fibrose Pulmonar/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Células Epiteliais Alveolares/patologia , Animais , Bleomicina/farmacologia , Células da Medula Óssea/patologia , Linhagem Celular , Feminino , Células-Tronco Mesenquimais/patologia , Camundongos , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/patologia , Fibrose Pulmonar/prevenção & controle
20.
Chin Med J (Engl) ; 132(8): 948-956, 2019 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-30958437

RESUMO

BACKGROUND: Cervical cancer has the fourth highest incidence and mortality rate of all cancers in women worldwide; it seriously harms their physical and mental health. The aim of this study was to observe the roles and preliminary mechanism of Taurine (Tau)-induced apoptosis in cervical cancer cells. METHODS: Cells from the human cervical cancer cell line SiHa were transfected with the recombinant plasmid pEGFP-N1-MST1 (mammalian sterile 20-like kinase 1); then, the cell proliferation activity was analyzed by the MTT assay, cell apoptosis by flow cytometry, and the related protein levels by Western blotting. RESULTS: Tau inhibited the proliferation of SiHa cells and induced apoptosis in these cells (the apoptotic rate was 21.95% in the Tau 160 mmol/L group and 30% in the Tau 320 mmol/L group), upregulated the expression of the MST1 (control, 0.53; Tau 40-320 mmol/L groups, 0.84-1.45) and Bax (control, 0.45; Tau 40-320 mmol/L groups, 0.64-1.51) proteins (P < 0.01), and downregulated the expression of Bcl-2 (control, 1.28, Tau 40-320 mmol/L groups, 0.93-0.47) (P < 0.01). The overexpression of MST1 promoted the apoptosis of SiHa cells, enhanced the apoptosis-inductive effects of Tau (P < 0.01), upregulated the expression of the proapoptotic proteins p73, p53, PUMA (p53 upregulated modulator of apoptosis), and caspase-3, and promoted the phosphorylation of YAP (Yes-associated protein). CONCLUSIONS: Tau inhibited the proliferation and induced the apoptosis of cervical cancer SiHa cells. The MST1 protein plays an important role in the Tau-induced apoptosis of cervical cancer cells.


Assuntos
Taurina/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Feminino , Fator de Crescimento de Hepatócito/metabolismo , Humanos , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Taurina/efeitos dos fármacos , Neoplasias do Colo do Útero/metabolismo , Proteína X Associada a bcl-2/metabolismo
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