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1.
Int J Biol Sci ; 18(7): 2914-2931, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35541901

RESUMO

Background: Control of ER-mitochondrial Ca2+ fluxes is a critical checkpoint to determine cell fate under stress. The 75-kDa glucose-regulated protein (GRP75) is a key tether protein facilitating mitochondria-associated ER membrane (MAM) formation through the IP3R-GRP75-VDAC1 complex. Although GRP75 contributes to cisplatin (CP)-resistance of ovarian cancer (OC), the underlying mechanisms are not clear. Methods: CP-resistant and -sensitive OC cell lines with GRP75 stable modulation were established. Confocal, PLA, co-IP, and TEM analysis were utilized to detect MAM integrity. Live cell Ca2+ imaging, intracellular ATP, ROS, and NAD+ assays were utilized to investigate ER-to-mitochondrial Ca2+ transfer and mitochondrial bioenergetics. Western blot, flow cytometry, CCK-8, Δψm, and mPTP assays were utilized to examine apoptotic cell death. Bioinformatics, patient's specimens, and immunohistochemistry were conducted to obtain the clinical relevance for GRP75-facilitated MAM formation. Results: GRP75-faciliated MAM formation was enriched in CP-resistant OC cells. CP-exposure only increased MAM formation in CP-sensitive OC cells, and enrichment of GRP75 and VDAC1 at MAMs is indispensable to CP-resistance. Diminishing MAM integrity by GRP75-deficiency reduced ER-to-mitochondria Ca2+ transfer, accelerated CP-induced mitochondrial dysfunction, provoked catastrophic ROS, and enhanced CP-triggered apoptotic cell death in OC cells. Clinical investigations confirmed the enrichment of GRP75-faciliated MAM formation in relapsed OC patients, and such enrichment was associated with the CP-resistance phenotype. Conclusion: GRP75-overexpression confers CP-resistance by distinctively managing MAM-facilitated Ca2+ fluxes and the pro-survival ROS signal, whereas GRP75-deficiency induces cell death via bioenergetic crisis and apoptotic ROS accumulation in OC cells. Our results show that GRP75-faciliated MAM formation is a potential target to overcome CP-resistance of OC.


Assuntos
Cisplatino , Proteínas de Choque Térmico HSP70 , Proteínas Mitocondriais , Neoplasias Ovarianas , Cálcio/metabolismo , Cisplatino/farmacologia , Resistencia a Medicamentos Antineoplásicos , Feminino , Proteínas de Choque Térmico HSP70/metabolismo , Humanos , Proteínas de Membrana/metabolismo , Mitocôndrias/metabolismo , Proteínas Mitocondriais/metabolismo , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo , Espécies Reativas de Oxigênio/metabolismo
2.
Infect Immun ; 89(12): e0029721, 2021 11 16.
Artigo em Inglês | MEDLINE | ID: mdl-34491790

RESUMO

Human cystic echinococcosis, caused by the larval stage of Echinococcus granulosus sensu lato, has been reported a near-cosmopolitan zoonotic disease. Various infiltrating immune cells gather around the lesion and produce a lesion microenvironment; however, cellular composition and heterogeneity in hepatic cystic echinococcosis lesion microenvironments are incompletely understood. Here, 81,865 immune cells isolated from peripheral blood, perilesion liver tissue, and adjacent normal liver tissue from four cystic echinococcosis patients were profiled using single-cell RNA sequencing. We identified 23 discrete cell populations and found distinct differences in infiltrating immune cells between tissue environments. Despite the significant similarity between perilesion and adjacent normal liver tissue-resident immune cells, the cellular proportions of type 2 innate lymphoid cells (ILC2s) and plasmacytoid dendritic cells (pDCs) were higher in perilesion liver tissue. Interestingly, the immunosuppressive gene NFKBIA was upregulated in these cells. Seven subsets of CD4+ T cell populations were found, and there were more regulatory-CD4+ T cells (Treg-CD4+) and Th2-CD4+ T cells in perilesion tissue than in adjacent normal tissue. There was close contact between CD4+ T cells and ILC2s and pDCs, which caused upregulation of genes related to positive immune activity in adjacent normal liver tissue. However, expression of genes related to immunosuppression, especially the immune inhibitory checkpoint gene NKG2A/HLA-E, was obviously higher in perilesion tissue, suggesting that cellular interaction resulted in an inhibitory microenvironment in the cystic echinococcosis (CE) lesion. This work offers new insights into the transcriptional heterogeneity of infiltrating immune cells in hepatic cystic echinococcosis lesion microenvironments at a single-cell level and provides potential target signatures for diagnosis and immunotherapies.


Assuntos
Microambiente Celular , Suscetibilidade a Doenças , Equinococose Hepática/etiologia , Equinococose Hepática/patologia , Interações Hospedeiro-Parasita , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Microambiente Celular/imunologia , Células Dendríticas , Sequenciamento de Nucleotídeos em Larga Escala , Interações Hospedeiro-Parasita/genética , Interações Hospedeiro-Parasita/imunologia , Humanos , Imunidade Inata , Linfócitos/imunologia , Linfócitos/metabolismo , Linfócitos/patologia , Análise de Célula Única
3.
Hepatology ; 74(6): 3376-3393, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34192365

RESUMO

BACKGROUND AND AIMS: Alveolar echinococcosis (AE) is a lethal helminthic liver disease caused by persistent infection with Echinococcus multilocularis. Although more attention has been paid to the immunotolerance of T cells caused by E. multilocularis infection, the role of natural killer (NK) cell, a critical player in liver immunity, is seldom studied. APPROACH AND RESULTS: Here, we observed that NK cells from the blood and closed liver tissue (CLT) of AE patients expressed a higher level of inhibitory receptor TIGIT and were functionally exhausted with a lower expression of granzyme B, perforin, interferon-gamma (IFN-γ), and TNF-α. Addition of anti-TIGIT (T-cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain) monoclonal antibody into AE patients' peripheral blood mononuclear cell culture significantly enhanced the synthesis of IFN-γ and TNF-α by NK cells, indicating the reversion of exhausted NK cells by TIGIT blockade. In the mouse model of E. multilocularis infection, liver and splenic TIGIT+ NK cells progressively increased dependent of infection dosage and timing and were less activated and less degranulated with lower cytokine secretion. Furthermore, TIGIT deficiency or blockade in vivo inhibited liver metacestode growth, reduced liver injury, and increased the level of IFN-γ produced by liver NK cells. Interestingly, NK cells from mice with persistent chronic infection expressed a higher level of TIGIT compared to self-healing mice. To look further into the mechanisms, more regulatory CD56bright and murine CD49a+ NK cells with higher TIGIT expression existed in livers of AE patients and mice infected with E. multilocularis, respectively. They coexpressed higher surface programmed death ligand 1 and secreted more IL-10, two strong inducers to mediate the functional exhaustion of NK cells. CONCLUSIONS: Our results indicate that inhibitory receptor TIGIT is involved in NK cell exhaustion and immune escape from E. multilocularis infection.


Assuntos
Equinococose/microbiologia , Receptores Imunológicos/metabolismo , Animais , Modelos Animais de Doenças , Equinococose/imunologia , Equinococose/metabolismo , Humanos , Células Matadoras Naturais/patologia , Camundongos
4.
J Int Med Res ; 48(8): 300060520946536, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32809866

RESUMO

Brenner tumor is a rare neoplasm of the vagina. This tumor is diagnosed according to the criteria of ovarian tumors. We report here a 64-year-old postmenopausal woman with a 2.0-cm sessile vaginal polyp for 9 years. Microscopic examination showed unusual features of no gland appearing in the tumor, but the other two characteristic components of transitional islands and dense fibrous stroma were observed. The tumor was diagnosed as a vaginal Brenner tumor on the basis of the definition proposed by the World Health Organization classification of female reproductive organ tumors. In our case, part of the epithelial nests of the Brenner tumor showed basaloid cell differentiation with peripheral palisading, and irregular papillary hyperplasia was observed around the epithelial nests similar to a borderline tumor. However, no mitotic activity or nuclear atypia was present in either the epithelial or stromal components. The presence of epithelial nests requires attention in the medical history of the patient. Our patient did not have a history of primary urothelial carcinoma. Our patient's benign vaginal Brenner tumor with different morphological characteristics supports the current notion that Walthard nests might act as possible precursor lesions.


Assuntos
Tumor de Brenner , Carcinoma de Células de Transição , Neoplasias Ovarianas , Tumor de Brenner/diagnóstico , Tumor de Brenner/cirurgia , Feminino , Humanos , Pessoa de Meia-Idade
5.
World J Clin Cases ; 8(12): 2448-2463, 2020 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-32607322

RESUMO

Chemokine (C-X-C motif) receptor 7 (CXCR7), recently termed ACKR3, belongs to the G protein-coupled cell surface receptor family, binds to stromal cell-derived factor-1 [SDF-1, or chemokine (C-X-C motif) ligand 12] or chemokine (C-X-C motif) ligand 11, and is the most common chemokine receptor expressed in a variety of cancer cells. SDF-1 binds to its receptor chemokine (C-X-C motif) receptor 4 (CXCR4) and regulates cell proliferation, survival, angiogenesis and migration. In recent years, another new receptor for SDF-1, CXCR7, has been discovered, and CXCR7 has also been found to be expressed in a variety of tumor cells and tumor-related vascular endothelial cells. Many studies have shown that CXCR7 can promote the growth and metastasis of a variety of malignant tumor cells. Unlike CXCR4, CXCR7 exhibits a slight modification in the DRYLAIV motif and does not induce intracellular Ca2+ release following ligand binding, which is essential for recruiting and activating G proteins. CXCR7 is generally thought to work in three ways: (1) Recruiting ß-arrestin 2; (2) Heterodimerizing with CXCR4; and (3) Acting as a "scavenger" of SDF-1, thus lowering the level of SDF-1 to weaken the activity of CXCR4. In the present review, the expression and role of CXCR7, as well as its prognosis in cancers of the digestive system, were investigated.

6.
Medicine (Baltimore) ; 99(26): e20854, 2020 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-32590784

RESUMO

INTRODUCTION: Although primary hepatic neuroendocrine carcinomas, whose prognostic mechanisms remain unclear, are rare, coexistence of neuroendocrine carcinomas and other tumors is rarer. In this report, we describe a unique case of coexistence between primary hepatic neuroendocrine carcinoma and a distal cholangiocarcinoma in the pancreas. PATIENT CONCERNS: A 64-year-old woman with a history of diabetes, but none of hepatitis, was admitted to hospital because of intermittent epigastric distension and pain discomfort for more than 1 month aggravated 1 day. A contrast-enhanced computed tomography (CT) scan of the upper abdomen and abdominal magnetic resonance imaging (MRI) revealed a thickening of the bile duct wall in the middle and lower segment of common bile duct and the corresponding lumen is narrow and low-density tumors with ring enhancement (1.83 cm × 1.9 cm) in lobi hepatis dexte. DIAGNOSIS: Primary neuroendocrine carcinoma of the liver was diagnosed to be coexisting with a distal cholangiocarcinoma, which had invaded the pancreas. Immunohistochemical examination revealed that the neoplastic cells strongly expressed chromogranin A, synaptophysin, and CD56 proteins. The tumor cells did not express HepPar-1, glypican-3, S-100, CK7, and CK19 in the liver tumor. A distal bile duct in pancreatic tissues shows the characteristics of typical bile duct carcinoma, as an invasion of carcinoma is also seen in the pancreatic tissues. Gastrointestinal endoscopy, chest and abdominal CT, abdominal MRI, and positron emission tomography (PET)-CT were used to exclude metastatic neuroendocrine tumors of the liver. INTERVENTIONS: Resection of the pancreas-duodenum, the right anterior lobe of the liver, and regional lymph nodes was performed in patients. OUTCOMES: The patient had survived for 5 months after the operation. CONCLUSION: A unique case of a coexistence of primary hepatic neuroendocrine carcinoma and a distal cholangiocarcinoma, which had invaded the pancreas. No treatment guidelines are established for the treatment of the unique case.


Assuntos
Carcinoma Neuroendócrino/diagnóstico , Colangiocarcinoma/diagnóstico , Fígado/anormalidades , Antígeno CD56/análise , Antígeno CD56/sangue , Carcinoma Neuroendócrino/patologia , Colangiocarcinoma/patologia , Cromogranina A/análise , Cromogranina A/sangue , Feminino , Humanos , Imuno-Histoquímica/métodos , Fígado/patologia , Fígado/fisiopatologia , Pessoa de Meia-Idade , Prognóstico , Sinaptofisina/análise , Sinaptofisina/sangue , Tomografia Computadorizada por Raios X/métodos
7.
Infect Genet Evol ; 80: 104198, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-31958516

RESUMO

Single-cell sequencing (SCS) is a next-generation sequencing method that is mainly used to analyze differences in genetic and protein information between cells, to obtain genetic information on microorganisms that are difficult to cultivate at a single-cell level and to better understand their specific roles in the microenvironment. By sequencing the whole genome, transcriptome and epigenome of a single cell, the complex heterogeneous mechanisms involved in disease occurrence and progression can be revealed, further improving disease diagnosis, prognosis prediction and monitoring of the therapeutic effects of drugs. In this study, we mainly summarized the methods and application fields of SCS, which may provide potential references for its future clinical applications, including the analysis of embryonic and organ development, the immune system, cancer progression, and parasitic and infectious diseases as well as stem cell research, antibody screening, and therapeutic research and development.


Assuntos
Sequenciamento de Nucleotídeos em Larga Escala/métodos , Análise de Célula Única/métodos , Epigênese Genética , Genoma , Genômica/métodos , Humanos , Separação Imunomagnética/métodos , Microdissecção/métodos , Microfluídica/instrumentação , Microfluídica/métodos , Especificidade de Órgãos , Análise de Sequência de DNA/métodos , Especificidade da Espécie , Transcriptoma
8.
Hepatology ; 71(4): 1297-1315, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31410870

RESUMO

BACKGROUND AND AIMS: The cestode Echinococcus multilocularis infection, a serious health problem worldwide, causes alveolar echinococcosis (AE), a tumor-like disease predominantly located in the liver and able to spread to any organs. Until now, there have been few studies that explore how T-cell exhaustion contributes to the parasite's escape from immune attack and how it might be reversed. APPROACH AND RESULTS: In this study, we found that liver T-cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT) expression was significantly enhanced and positively correlated with lesion activity in AE patients. High TIGIT expression in both liver-infiltrating and blood T cells was associated with their functional exhaustion, and its ligand CD155 was highly expressed by hepatocytes surrounding the infiltrating lymphocytes. In co-culture experiments using human blood T cells and hepatic cell line HL-7702, CD155 induced functional impairment of TIGIT+ T cells, and in vitro blockade with TIGIT antibody restored the function of AE patients' T cells. Similar TIGIT-related functional exhaustion of hepatic T cells and an abundant CD155 expression on hepatocytes were observed in E. multilocularis-infected mice. Importantly, in vivo blocking TIGIT prevented T-cell exhaustion and inhibited disease progression in E. multilocularis-infected mice. Mechanistically, CD4+ T cells were totally and CD8+ T cells partially required for anti-TIGIT-induced regression of parasite growth in mice. CONCLUSIONS: This study demonstrates that E. multilocularis can induce T-cell exhaustion through inhibitory receptor TIGIT, and that blocking this checkpoint may reverse the functional impairment of T cells and represent a possible approach to immunotherapy against AE.


Assuntos
Anticorpos Monoclonais/uso terapêutico , Linfócitos T CD8-Positivos/imunologia , Equinococose Hepática/terapia , Equinococose/terapia , Receptores Imunológicos/antagonistas & inibidores , Animais , Linhagem Celular , Modelos Animais de Doenças , Equinococose/imunologia , Equinococose Hepática/imunologia , Feminino , Humanos , Imunoterapia/métodos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Receptores Imunológicos/imunologia , Receptores Virais
9.
Front Immunol ; 11: 600635, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33488594

RESUMO

Echinococcus multilocularis larvae, predominantly located in the liver, cause a tumor-like parasitic disease, alveolar echinococcosis (AE), that is characterized by increased infiltration of various immune cells, including macrophages, around the lesion that produces an "immunosuppressive" microenvironment, favoring its persistent infection. However, the role of hepatic macrophages in the host defense against E. multilocularis infection remains poorly defined. Using human liver tissues from patients with AE and a hepatic experimental mouse model of E. multilocularis, we investigated the phenotype and function of hepatic macrophages during the parasite infection. In the present study, we found that a large number of CD68+ macrophages accumulated around the metacestode lesion in the liver of human AE samples and that both S100A9+ proinflammatory (M1 phenotype) and CD163+ anti-inflammatory (M2 phenotype) macrophages were significantly higher in close liver tissue (CLT) than in distant liver tissue (DLT), whereas M2 macrophages represent the dominant macrophage population. Furthermore, E. multilocularis-infected mice exhibited a massive increase in macrophage (F4/80+) infiltration in the liver as early as day 5, and the infiltrated macrophages were mainly monocyte-derived macrophages (CD11bhi F4/80int MoMFs) that preferentially differentiated into the M1 phenotype (iNOS+) at the early stage of E. multilocularis infection and then polarized to anti-inflammatory macrophages of the M2 phenotype (CD206+) at the chronic stage of infection. We further showed that elimination of macrophages by treatment of mice with clodronate-liposomes before E. multilocularis infection impaired worm expulsion and was accompanied by a reduction in liver fibrosis, yielding a high parasite burden. These results suggest that hepatic macrophages may play a dual role in the establishment and development of E. multilocularis metacestodes in which early larvae clearance is promoted by M1 macrophages while persistent metacestode infection is favored by M2 macrophages.


Assuntos
Equinococose , Echinococcus multilocularis/imunologia , Estágios do Ciclo de Vida/imunologia , Fígado , Macrófagos , Animais , Equinococose/imunologia , Equinococose/parasitologia , Equinococose/patologia , Feminino , Humanos , Fígado/imunologia , Fígado/parasitologia , Fígado/patologia , Macrófagos/imunologia , Macrófagos/parasitologia , Macrófagos/patologia , Camundongos
10.
Sci Rep ; 9(1): 19612, 2019 12 23.
Artigo em Inglês | MEDLINE | ID: mdl-31873157

RESUMO

The local immune mechanisms responsible for the establishment and development of Echinococcus granulosus sensu stricto infection in the liver, have been little explored. We developed a suitable experimental model that mimics naturally infected livers using portal injection of protoscoleces. Opposite to Echinococcus multilocularis infection which is dose-dependent, fully mature hydatid cysts can be established in the liver whatever the injection dose; although most of the infection sites were seen at the establishment phase as inflammatory granulomas associated with fibrosis, they never matured into cysts. At the establishment phase, a strong immune response was composed of T and B cells, with T1-type, T2-type cells and cytokines and IL-10-secreting CD8+ T cells in the liver. At the established phase, results suggested a local production of antibodies by B cells, and an involvement of NK and NKT cells. Infection outcome and local immune response in the liver, were different in the mouse models of Echinococcus granulosus sensu stricto and Echinococcus multilocularis respectively; however, only early specificities at the microenvironment level might explain the major differences found between the lesions induced by the two species. Our quantitative experimental model appears fully appropriate to further study this microenvironment and its relationship with each cestode species.


Assuntos
Equinococose , Echinococcus granulosus/imunologia , Hepatopatias Parasitárias , Fígado , Animais , Linfócitos B/imunologia , Linfócitos B/patologia , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/patologia , Modelos Animais de Doenças , Equinococose/imunologia , Equinococose/patologia , Feminino , Interleucina-10/imunologia , Fígado/imunologia , Fígado/parasitologia , Fígado/patologia , Hepatopatias Parasitárias/imunologia , Hepatopatias Parasitárias/parasitologia , Hepatopatias Parasitárias/patologia , Camundongos , Células Th1/imunologia , Células Th1/patologia , Células Th2/imunologia , Células Th2/patologia
11.
Parasit Vectors ; 12(1): 542, 2019 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-31727141

RESUMO

BACKGROUND: Larvae of Echinococcus granulosus (sensu lato) dwell in host organs for a long time but elicit only a mild inflammatory response, which indicates that the resolution of host inflammation is necessary for parasite survival. The recruitment of alternatively activated macrophages (AAMs) has been observed in a variety of helminth infections, and emerging evidence indicates that AAMs are critical for the resolution of inflammation. However, whether AAMs can be induced by E. granulosus (s.l.) infection or thioredoxin peroxidase (TPx), one of the important molecules secreted by the parasite, remains unclear. METHODS: The activation status of peritoneal macrophages (PMs) derived from mice infected with E. granulosus (sensu stricto) was analyzed by evaluating the expression of phenotypic markers. PMs were then treated in vivo and in vitro with recombinant EgTPx (rEgTPx) and its variant (rvEgTPx) in combination with parasite excretory-secretory (ES) products, and the resulting activation of the PMs was evaluated by flow cytometry and real-time PCR. The phosphorylation levels of various molecules in the PI3K/AKT/mTOR pathway after parasite infection and antigen stimulation were also detected. RESULTS: The expression of AAM-related genes in PMs was preferentially induced after E. granulosus (s.s.) infection, and phenotypic differences in cell morphology were detected between PMs isolated from E. granulosus (s.s.)-infected mice and control mice. The administration of parasite ES products or rEgTPx induced the recruitment of AAMs to the peritoneum and a notable skewing of the ratio of PM subsets, and these effects are consistent with those obtained after E. granulosus (s.s.) infection. ES products or rEgTPx also induced PMs toward an AAM phenotype in vitro. Interestingly, this immunomodulatory property of rEgTPx was dependent on its antioxidant activity. In addition, the PI3K/AKT/mTOR pathway was activated after parasite infection and antigen stimulation, and the activation of this pathway was suppressed by pre-treatment with an AKT/mTOR inhibitor. CONCLUSIONS: This study demonstrates that E. granulosus (s.s.) infection and ES products, including EgTPx, can induce PM recruitment and alternative activation, at least in part, via the PI3K/AKT/mTOR pathway. These results suggest that EgTPx-induced AAMs might play a key role in the resolution of inflammation and thereby favour the establishment of hydatid cysts in the host.


Assuntos
Echinococcus granulosus/imunologia , Macrófagos Peritoneais/imunologia , Proteína Oncogênica v-akt/metabolismo , Peroxirredoxinas/imunologia , Fosfatidilinositol 3-Quinases/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Animais , Equinococose/parasitologia , Echinococcus granulosus/enzimologia , Feminino , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/parasitologia , Camundongos , Camundongos Endogâmicos BALB C , Peroxirredoxinas/farmacologia , Fenótipo , Fosforilação , Transdução de Sinais , Organismos Livres de Patógenos Específicos
12.
J Exp Clin Cancer Res ; 38(1): 471, 2019 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-31752956

RESUMO

BACKGROUND: Adhesion molecules distributed on the cell-surface depends upon their dynamic trafficking that plays an important role during cancer progression. ADP-ribosylation factor 6 (Arf6) is a master regulator of membrane trafficking. CD147, a tumor-related adhesive protein, can promote the invasion of liver cancer. However, the role of Arf6 in CD147 trafficking and its contribution to liver cancer progression remain unclear. METHODS: Stable liver cancer cell lines with Arf6 silencing and over-expression were established. Confocal imaging, flow cytometry, biotinylation and endomembrane isolation were used to detect CD147 uptake and recycling. GST-pull down, gelatin zymography, immunofluorescence, cell adhesion, aggregation and tight junction formation, Transwell migration, and invasion assays were used to examine the cellular phenotypes. GEPIA bioinformatics, patient's specimens and electronic records collection, and immunohistochemistry were performed to obtain the clinical relevance for Arf6-CD147 signaling. RESULTS: We found that the endocytic recycling of CD147 in liver cancer cells was controlled by Arf6 through concurrent Rab5 and Rab22 activation. Disruption of Arf6-mediated CD147 trafficking reduced the cell-matrix and cell-cell adhesion, weakened cell aggregation and junction stability, attenuated MMPs secretion and cytoskeleton reorganization, impaired HGF-stimulated Rac1 activation, and markedly decreased the migration and invasion of liver cancer cells. Moreover, high-expression of the Arf6-CD147 signaling components in HCC (hepatocellular carcinoma) was closely correlated with poor clinical outcome of patients. CONCLUSIONS: Our results revealed that Arf6-mediated CD147 endocytic recycling is required for the malignant phenotypes of liver cancer. The Arf6-driven signaling machinery provides excellent biomarkers or therapeutic targets for the prevention of liver cancer.


Assuntos
Fatores de Ribosilação do ADP/metabolismo , Basigina/metabolismo , Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/metabolismo , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Adesão Celular/fisiologia , Agregação Celular/fisiologia , Linhagem Celular Tumoral , Endocitose , Células Hep G2 , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Fenótipo , Regulação para Cima , Proteínas rab de Ligação ao GTP/metabolismo , Proteínas rab5 de Ligação ao GTP/metabolismo
13.
BMC Infect Dis ; 19(1): 792, 2019 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-31500589

RESUMO

BACKGROUND: Alveolar echinococcosis (AE) is caused by the larval stage of Echinococcus multilocularis (E. multilocularis), and considered as public health issue. Parasite-host immune interaction is pivotal during infection. As a subset of innate lymphoid cells, NK cells are known to play an important role during virus, bacteria, intra/extracellular parasitic infections and tumor progression. However, the possible role of NK cells in E. multilocularis infection in both human and murine is little known. Herein, the functional alteration of hepatic NK cells and their related molecules in E. multilocularis infected mice were studied. METHODS: 2000 protoscoleces (PSCs) were injected to C57BL/6 mice via the portal vein to establish secondary E. multilocularis infection. NK cells population and their related molecules (CD69, Ly49D, Ly49G2, Ly49H, Ly49I, NKG2A, NKG2D, granzyme B, IFN-γ, TNF-α) were assessed by using fluorescence-activated cell sorter (FACS) techniques and qRT-PCR. NK cell depletion was performed for further understanding the possible function of NK cells during infection. RESULTS: The total frequencies of NK cells and NK-derived IFN-γ production were significantly reduced at designated time points (2, 4, 12 weeks). The liver resident (CD49a+DX5-) NK cells are decreased at 4 weeks after inoculation and which is significantly lower than in control mice. Moreover, in vivo antibody-mediated NK cell depletion increased parasitic load and decreased peri-parasitic fibrosis. Expression of the inhibitory receptor NKG2A was negatively related to NK- derived IFN-γ secretion. CONCLUSIONS: Our study showed down regulates of NK cells and upper regulates of NKG2A expression on NK cells during E. multilocularis infection. Reduction of NK cell frequencies and increased NKG2A might result in low cytotoxic activity through decreased IFN-γ secretion in E. multilocularis infection. This result might be helpful to restore NK cell related immunity against E. multilocularis infection to treat alveolar echinococcosis.


Assuntos
Echinococcus multilocularis/fisiologia , Subfamília C de Receptores Semelhantes a Lectina de Células NK/metabolismo , Animais , Equinococose/microbiologia , Equinococose/patologia , Feminino , Tolerância Imunológica , Integrina alfa1/metabolismo , Interferon gama/metabolismo , Células Matadoras Naturais/citologia , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Fígado/imunologia , Fígado/patologia , Camundongos , Camundongos Endogâmicos C57BL , Subfamília C de Receptores Semelhantes a Lectina de Células NK/genética , Regulação para Cima/efeitos dos fármacos
14.
BMC Musculoskelet Disord ; 20(1): 356, 2019 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-31376836

RESUMO

BACKGROUND: Fibrous hamartoma of infancy(FHI) is a rare benign lesion most frequently occurring within the first year of life. So far, just over 200 cases have been reported in the English literature, in which the radiologic findings of FHI have not been fully described. Herein, 2 adult cases of FHI receiving treatment in our hospital and the published cases searched on PubMed are reviewed, with the emphasis on the discussion of the spectrum of MR findings and their histologic correlation. CASE PRESENTATION: We present two adult cases who aged 47 years and 19 years with slow growing masses beginning from their childhood in the posterior craniocervical area. On CT and MR imaging, the tumours showed as the superficially located lesions with ill-defined margins that involved the subcutaneous layer and its underlying muscles. The size of the lesions were 21.3 × 16.7 × 16 cm in case 1 and 20.2 × 19.3 × 13.6 cm in case 2. The tumours demonstrated heterogeneous intensities/signals with the adipose tissue presenting as the disperse strands or small focus of fatty intensity/signal. Parallel or whirling appearance, and dilated vessels were delineated in the cases. Contrast enhancement was administered in case 1 and marked enhancement was found. CONCLUSIONS: The usually observed manifestation of FHI on CT and/or MR imaging is the strands of adipose/fibrous intensities traversing the lesions, with the characteristic parallel or whirling appearance in some cases. The tumours with ill-defined margins have the tendency to involve the underlying muscles. Some fibroblastic and adipocytic tumours should be ruled out in differential diagnosis.


Assuntos
Hamartoma/diagnóstico por imagem , Tela Subcutânea/patologia , Diagnóstico Diferencial , Feminino , Fibrose , Hamartoma/patologia , Hamartoma/cirurgia , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Pescoço , Tela Subcutânea/diagnóstico por imagem , Tela Subcutânea/cirurgia , Tomografia Computadorizada por Raios X , Resultado do Tratamento , Adulto Jovem
15.
Sci Rep ; 9(1): 3424, 2019 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-30809024

RESUMO

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.

16.
J Histochem Cytochem ; 67(5): 299-308, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30633620

RESUMO

Chemokines and their receptors have been proposed to play important roles in tumor progression and metastasis. To investigate their roles in the progression of primary and metastatic malignant liver tumors and their prognosis, we compared expression profiles of CXCL12/CXCR4, CCL20/CCR6, and CCL21/CCR7 in hepatocellular carcinoma (HCC) and colorectal liver metastases (CRLM). Immunohistochemistry was used to analyze the expression levels of the chemokine/chemokine receptor pairs in 29 HCC and 11 CRLM specimens and adjacent non-cancerous tissues, and correlations with clinicopathological variables and overall survival were determined. CCL20/CCR6 expression was higher in HCC than in adjacent non-cancerous tissues. High CCR6 expression in HCC was negatively associated with 5-year survival rate and was an independent prognostic factor for overall survival of HCC patients, whereas differences were not observed between CRLM and adjacent tissues. Furthermore, significantly higher expression of CCL21/CCR7 was found in CRLM than in HCC. In summary, the CCL20/CCR6 axis was elevated in HCC but not in CRLM, whereas the CCL21/CCR7 axis was elevated in CRLM but not in HCC.


Assuntos
Carcinoma Hepatocelular/diagnóstico , Quimiocinas/análise , Neoplasias Colorretais/patologia , Neoplasias Hepáticas/diagnóstico , Fígado/patologia , Receptores de Quimiocinas/análise , Adulto , Idoso , Biomarcadores Tumorais/análise , Carcinoma Hepatocelular/secundário , Quimiocina CCL20/análise , Quimiocina CCL21/análise , Feminino , Humanos , Neoplasias Hepáticas/secundário , Masculino , Pessoa de Meia-Idade , Prognóstico , Receptores CCR6/análise , Receptores CCR7/análise
17.
Artigo em Inglês | MEDLINE | ID: mdl-30348669

RESUMO

Cystic echinococcosis is a zoonosis caused by the larval stage of Echinococcus granulosus sensu lato There is an urgent need to develop new drugs for the treatment of this disease. In this study, we identified two new members of mitogen-activated protein kinase (MAPK) cascades, MKK3/6 and MEK1/2 homologs (termed EgMKK1 and EgMKK2, respectively), from E. granulosus sensu stricto Both EgMKK1 and EgMKK2 were expressed at the larval stages. As shown by yeast two-hybrid and coimmunoprecipitation analyses, EgMKK1 interacted with the previously identified Egp38 protein but not with EgERK. EgMKK2, on the other hand, interacted with EgERK. In addition, EgMKK1 and EgMKK2 displayed kinase activity toward the substrate myelin basic protein. When sorafenib tosylate, PD184352, or U0126-ethanol (EtOH) was added to the medium for in vitro culture of E. granulosus protoscoleces (PSCs) or cysts, an inhibitory and cytolytic effect was observed via suppressed phosphorylation of EgMKKs and EgERK. Nonviability of PSCs treated with sorafenib tosylate or U0126-EtOH, and not with PD184352, was confirmed through bioassays, i.e., inoculation of treated and untreated protoscoleces into mice. In vivo treatment of E. granulosus sensu stricto-infected mice with sorafenib tosylate or U0126-EtOH for 4 weeks demonstrated a reduction in parasite weight, but the results did not show a significant difference. In conclusion, the MAPK cascades were identified as new targets for drug development, and E. granulosus was efficiently inhibited by their inhibitors in vitro The translation of these findings into in vivo efficacy requires further adjustment of treatment regimens using sorafenib tosylate or, possibly, other kinase inhibitors.


Assuntos
Benzamidas/farmacologia , Butadienos/farmacologia , Equinococose/tratamento farmacológico , Echinococcus granulosus/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Nitrilas/farmacologia , Sorafenibe/farmacologia , Animais , Equinococose/parasitologia , Equinococose/patologia , Echinococcus granulosus/metabolismo , MAP Quinase Quinase 1/antagonistas & inibidores , MAP Quinase Quinase 2/antagonistas & inibidores , MAP Quinase Quinase 3/antagonistas & inibidores , MAP Quinase Quinase 6/antagonistas & inibidores , Camundongos , Camundongos Endogâmicos BALB C , Fosforilação/efeitos dos fármacos
18.
Parasite ; 25: 57, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30474598

RESUMO

Cystic echinococcosis (CE) is a cosmopolitan parasitic disease caused by infection with the larval stage of Echinococcus granulosus sensu lato. Thioredoxin peroxidase (TPx) may play an essential role in the antioxidant defence system of E. granulosus s.l. as neither catalase nor glutathione peroxidase activities have been detected in the parasite. However, it is not known whether TPx affects the survival and growth of E. granulosus s.l. during development. In this study, three fragments of siRNA specific for EgTPx (siRNA-1/2/3) were designed and transfected into protoscoleces of E. granulosus sensu stricto by electroporation. Quantitative real-time PCR and Western blotting analysis showed that siRNA-3 significantly reduced the expression of EgTPx. Coincidentally, knockdown of EgTPx expression in protoscoleces with siRNA-3 significantly reduced the viability of the parasite under oxidative stress induced by 0.6 mM H2O2. In vitro culture studies showed that protoscoleces treated with siRNA-3 reduced pre-microcyst formation. In vivo experiments showed that injecting mice intraperitoneally with protoscoleces treated with siRNA-3 resulted in a significant reduction in the number, size and weight of CE cysts compared with those of control animals. Silencing of EgTPx led to the impairment of growth of E. granulosus s.s. both in vitro and in vivo, indicating that EgTPx is an important factor for protoscoleces survival and plays an important role in the antioxidant defence against the host during development.


Assuntos
Echinococcus granulosus/enzimologia , Echinococcus granulosus/fisiologia , Peroxirredoxinas/deficiência , Peroxirredoxinas/genética , Animais , Western Blotting , Equinococose/parasitologia , Echinococcus granulosus/genética , Echinococcus granulosus/crescimento & desenvolvimento , Eletroporação , Feminino , Inativação Gênica , Peróxido de Hidrogênio/farmacologia , Larva/fisiologia , Camundongos , Camundongos Endogâmicos BALB C , Interferência de RNA , RNA Interferente Pequeno/fisiologia , Reação em Cadeia da Polimerase em Tempo Real , Ovinos , Transfecção
19.
Am J Transl Res ; 10(9): 2949-2957, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30323881

RESUMO

The purpose of this study was to investigate the correlations between Gd-EOB-DTPA DCE-MRI biomarkers and histopathologic biomarkers of liver fibrosis progression in a rabbit model of liver fibrosis. Thirty-Six New Zealand white rabbits were randomly divided into control (n = 6) and liver fibrosis group (n = 30). Each rabbit in the liver fibrosis group received a weekly subcutaneous injection in the back comprising 50% carbon tetrachloride (CCl4) in oily solution. Control rabbits received subcutaneous injections with the same amount of normal saline solution instead. MR imaging was performed in control and fibrotic rabbits were conducted by MRI at week 0 (n = 36). The fibrotic rabbits were performed MR imaging on 6 weeks, 9 weeks, and 12 weeks after modeling of fibrosis. Before each MRI, peripheral blood was collected, and several biochemical testes are performed. The thirty-four rabbits completed this study. They were then divided into three subgroups according to fibrotic stage: no fibrosis (F0, n = 12), mild fibrosis (F1+F2, n = 14), and advanced fibrosis (F3+F4, n = 8). DCE-MRI measurements show increasing Ktrans and Ve while decreasing iACU90 with increasing fibrosis stage. The significant correlations were observed between mean Ktrans, Ve, iACU90 and percentage of the animal with mild liver fibrosis. For blood biomarkers, there were significant differences between F0 and F1+F2, and between F0 and F3+F4 in the serum levels of ALT (all P < 0.05), and TB (F0 vs. F1+F2, P = 0.004), while no differences were found between F1+F2 group and F3+F4 group (all P > 0.05). There were significant differences between F0 and F1+F2 (P = 0.02). Gd-EOB-DTPA DCE-MRI is a promising method for the noninvasive diagnosis and staging of liver fibrosis. Future studies to evaluate the effectiveness of these techniques in patients with liver fibrosis are warranted.

20.
Exp Ther Med ; 16(4): 3155-3160, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30214538

RESUMO

Alveolar echinococcosis (AE) is a prevalent epidemic in the northern hemisphere, especially in central Europe and western China. Serum diagnosis is important for patients with AE, especially during the first screening. The present study purified the recombinant Em18-GST (rEm18-GST), and detected its diagnostic performance in human alveolar echinococcosis patients of Xinjiang, China with immunoblotting (IB) and enzyme-linked immunosorbent assay (ELISA). Serum samples were collected from 50 patients with AE, 222 patients with cystic echinococcosis (CE), 158 patients with other unrelated infections and 106 healthy individuals. The IB results showed that serum samples of 47 patients with AE and 12 patients with CE were rEm18-positive. However, only one sample from patients with cancer showed a cross-reaction with rEm18 in IB. The overall sensitivity was 94%, and the total specificity was 96.58%. For the rEm18 results using ELISA, the sera of 46 patients with AE were positive, and the overall sensitivity was 92%. In conclusion, compared with imaging tools, including computed tomography, magnetic resonance imaging and positron emission tomography, rEm18 has considerable advantages for AE serodiagnosis.

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