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1.
Int J Mol Sci ; 22(4)2021 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-33672075

RESUMO

Positive experiences in early life may improve the capacity to cope with adulthood stress through epigenetic modification. We investigated whether an enriched environment (EE) in the postnatal period affected epigenetic changes in the p11 gene induced by chronic unpredictable stress (CUS) in adult C57BL/6J mice. EE was introduced for 5 weeks during postnatal days 21-55. After EE, the mice were subjected to CUS for 4 weeks. EE prevented depression-like behavior induced by adult CUS. EE prevented a decrease in p11 mRNA and histone H3 acetylation induced by CUS, with changes in the expression of histone deacetylase 5. Moreover, EE prevented changes in trimethylation of histone H3 lysine 4 (H3K4) and H3K27 induced by CUS. Furthermore, EE had positive effects on behavior and epigenetic alterations in adult mice without CUS. These results suggest that one of the underlying mechanisms of early-life EE may involve epigenetic modification of the hippocampal p11 gene promoter.


Assuntos
Anexina A2/genética , Depressão/sangue , Depressão/prevenção & controle , Epigênese Genética , Expressão Gênica , Abrigo para Animais , Proteínas S100/genética , Estresse Fisiológico , Acetilação , Animais , Corticosterona/sangue , Hipocampo/metabolismo , Histona Desacetilases/metabolismo , Histonas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Regiões Promotoras Genéticas/genética , RNA Mensageiro/genética
2.
3.
Phys Chem Chem Phys ; 23(5): 3519-3530, 2021 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-33514968

RESUMO

Glycosaminoglycans (GAGs) are anionic, periodic, linear polysaccharides which are composed of periodic disaccharide units. They play a vital role in many biological processes ongoing in the extracellular matrix. In terms of computational approaches, GAGs are very challenging molecules due to their high flexibility, periodicity, predominantly electrostatic-driven nature of interactions with their protein counterparts and potential multipose binding. Furthermore, the molecular mechanisms underlying GAG-mediated interactions are not fully known yet, and experimental techniques alone are not always sufficient to gain insights into them. The aim of this study was to characterize protein-ion-GAG complexes for the systems where ions are directly involved in GAG binding. Molecular docking, molecular dynamics and free energy calculation approaches were applied to model and rigorously analyse the interactions between annexins (II and V), calcium ions (Ca2+) and heparin (HP). The computational data were examined and discussed in the context of the structural data previously reported by the crystallographic studies. The computational results confirm that the presence of Ca2+ has a tremendous impact on the annexin-HP binding site. This study provides a general computational pipeline to discover the complexity of protein-GAG interactions and helps to understand the role of ions involved at the atomic level. The limitations of the applied protocols are described and discussed pointing at the challenges persisting in the state-of-the-art in silico tools to study protein-ion-GAG systems.


Assuntos
Anexina A2/metabolismo , Anexina A5/metabolismo , Cálcio/metabolismo , Heparina/metabolismo , Animais , Anexina A2/química , Anexina A5/química , Cálcio/química , Heparina/química , Humanos , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Ligação Proteica , Ratos , Termodinâmica
4.
Biochim Biophys Acta Biomembr ; 1863(1): 183451, 2021 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-32835655

RESUMO

AIIt, a heterotetramer of S100A10 (P11) and Annexin A2, plays a key role in calcium dependent, membrane associations with a variety of proteins. We previously showed that AIIt interacts with the short cytoplasmic domain (12 amino acids) of CEACAM1 (CEACAM1-SF). Since the cytoplasmic domains of CEACAM1 help regulate the formation of cis- or trans-dimers at the cell membrane, we investigated the possible role of their association with AIIt in this process. Using NMR and molecular dynamics, we show that AIIt and its pseudoheterodimer interacts with two molecules of short cytoplasmic domain isoform peptides, and that interaction depends on the binding motif 454-Phe-Gly-Lys-Thr-457 where Phe-454 binds in a hydrophobic pocket of AIIt, the null mutation Phe454Ala reduces binding by 2.5 fold, and the pseudophosphorylation mutant Thr457Glu reduces binding by three fold. Since these two residues in CEACAM1-SF were also found to play a role in the binding of calmodulin and G-actin at the membrane, we hypothesize a sequential set of three interactions are responsible for regulation of cis- to trans-dimerization of CEACAM1. The hydrophobic binding pocket in AIIt corresponds to a previously identified binding pocket for a peptide found in SMARCA3 and AHNAK, suggesting a conserved functional motif in AIIt allowing multiple proteins to reversibly interact with integral membrane proteins in a calcium dependent manner.


Assuntos
Anexina A2/química , Antígenos CD/química , Moléculas de Adesão Celular/química , Simulação de Dinâmica Molecular , Complexos Multiproteicos/química , Ressonância Magnética Nuclear Biomolecular , Proteínas S100/química , Humanos , Domínios Proteicos , Multimerização Proteica
5.
Sci Rep ; 10(1): 22206, 2020 12 17.
Artigo em Inglês | MEDLINE | ID: mdl-33335142

RESUMO

Collagen type VI (COL6) deposition occurs in various glomerular diseases, causing serious pathological damage like nodular lesions. However, the mechanisms underlying the deposition of COL6 remain unclear. In renal biopsy samples, immunohistochemical analyses revealed that COL6 and phosphorylated histone H2AX (γ-H2AX), a DNA damage marker, were detected mainly in diabetic nodular glomerulosclerosis, in which the γ-H2AX-positive area was identified as the independent factor significantly associated with the COL6-positive area (ß: 0.539, t = 2.668). In in vitro studies, COL6 secretion from human renal glomerular endothelial cells (HRGECs) was assessed by measuring the decrease in the cytoplasmic COL6-positive cells and an increase in the amount of COL6 in the culture medium. Mitomycin C (MMc) treatment of HRGECs increased the number of γ-H2AX-positive cells and COL6 secretion, which were suppressed by a specific inhibitor of ataxia telangiectasia and Rad3-related (ATR). MMc-induced COL6 secretion was also suppressed by Annexin A2 (ANXA2) siRNA transfection. Moreover, the inhibition of ATR activity did not induce any extra suppression in the MMc-induced COL6 secretion by ANXA2 siRNA transfected cells. These results confirm that nodular glomerulosclerosis partially results from DNA damage in the glomerulus and that DNA damage-induced COL6 secretion from HRGECs occurs through an ATR and ANXA2-mediated pathway.


Assuntos
Anexina A2/metabolismo , Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , Dano ao DNA , Nefropatias Diabéticas/etiologia , Nefropatias Diabéticas/metabolismo , Células Endoteliais/metabolismo , Glomérulos Renais/metabolismo , Glomérulos Renais/patologia , Adulto , Idoso , Biomarcadores , Biópsia , Colágeno Tipo VI/metabolismo , Nefropatias Diabéticas/patologia , Suscetibilidade a Doenças , Feminino , Imunofluorescência , Histonas/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Ligação Proteica
6.
Nat Commun ; 11(1): 4512, 2020 09 09.
Artigo em Inglês | MEDLINE | ID: mdl-32908147

RESUMO

Hydrogen peroxide (H2O2) is recognized to act as a signaling molecule. Peroxiredoxins (Prxs) have the ability to transfer H2O2-derived oxidizing equivalents to redox-regulated target proteins, thus facilitating the transmission of H2O2 signals. It has remained unclear how Prxs and their target proteins are brought together to allow for target-specific protein thiol oxidation. Addressing the specific case of Prx2-dependent STAT3 oxidation, we here show that the association of the two proteins occurs prior to Prx oxidation and depends on a scaffolding protein, the membrane chaperone annexin A2. Deletion or depletion of annexin A2 interrupts the transfer of oxidizing equivalents from Prx2 to STAT3, which is observed to take place on membranes. These findings support the notion that the Prx2-STAT3 redox relay is part of a highly organized membrane signaling domain.


Assuntos
Anexina A2/metabolismo , Peroxirredoxinas/metabolismo , Fator de Transcrição STAT3/metabolismo , Anexina A2/genética , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Dissulfetos/metabolismo , Células HEK293 , Humanos , Peróxido de Hidrogênio/metabolismo , Oxirredução , Ligação Proteica , Domínios Proteicos , Transdução de Sinais
7.
DNA Cell Biol ; 39(10): 1862-1871, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32845709

RESUMO

Sepsis is a life-threatening disorder and leads to organ dysfunction and death. Therefore, searching for more alternative biomarkers is of great significance for sepsis assessment and surveillance. In our study, the gene expression profiles of 163 samples from healthy controls and septic patients were analyzed and 8 gene co-expression modules were identified by constructing weighted gene co-expression network. The blue and yellow modules showed close correlations with the phenotypic trait "days postsepsis." Besides, differentially expressed genes (DEGs) over time in septic patients were screened using Short Time-series Expression Miner (STEM) program. The intersection of genes in the blue and yellow modules and DEGs, which were significantly enriched in "HTLV-1 infection" pathway, was analyzed with protein-protein interaction network. The logistic regression model based on these eight mRNAs was constructed to determine the type of the sample reliably. Eight vital genes CECR1, ANXA2, ELANE, CTSG, AZU1, PRTN3, LYZ, and DEFA4 presented high scores and may be associated with sepsis, which provided candidate biomarkers for sepsis.


Assuntos
Sepse/genética , Transcriptoma , Adenosina Desaminase/genética , Adenosina Desaminase/metabolismo , Anexina A2/genética , Anexina A2/metabolismo , Biomarcadores/metabolismo , Catepsina G/genética , Catepsina G/metabolismo , Redes Reguladoras de Genes , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Elastase de Leucócito/genética , Elastase de Leucócito/metabolismo , Mieloblastina/genética , Mieloblastina/metabolismo , Sepse/metabolismo , Sepse/patologia , Análise de Sobrevida , alfa-Defensinas/genética , alfa-Defensinas/metabolismo
8.
PLoS Negl Trop Dis ; 14(7): e0007960, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32687500

RESUMO

Intracerebral microhemorrhages (CMHs) are small foci of hemorrhages in the cerebrum. Acute infections induced by some intracellular pathogens, including rickettsia, can result in CMHs. Annexin a2 (ANXA2) has been documented to play a functional role during intracellular bacterial adhesion. Here we report that ANXA2-knockout (KO) mice are more susceptible to CMHs in response to rickettsia and Ebola virus infections, suggesting an essential role of ANXA2 in protecting vascular integrity during these intracellular pathogen infections. Proteomic analysis via mass spectrometry of whole brain lysates and brain-derived endosomes from ANXA2-KO and wild-type (WT) mice post-infection with R. australis revealed that a variety of significant proteins were differentially expressed, and the follow-up function enrichment analysis had identified several relevant cell-cell junction functions. Immunohistology study confirmed that both infected WT and infected ANXA2-KO mice were subjected to adherens junctional protein (VE-cadherin) damages. However, key blood-brain barrier (BBB) components, tight junctional proteins ZO-1 and occludin, were disorganized in the brains from R. australis-infected ANXA2-KO mice, but not those of infected WT mice. Similar ANXA2-KO dependent CMHs and fragments of ZO-1 and occludin were also observed in Ebola virus-infected ANXA2-KO mice, but not found in infected WT mice. Overall, our study revealed a novel role of ANXA2 in the formation of CMHs during R. australis and Ebola virus infections; and the underlying mechanism is relevant to the role of ANXA2-regulated tight junctions and its role in stabilizing the BBB in these deadly infections.


Assuntos
Anexina A2/metabolismo , Hemorragia Cerebral/metabolismo , Ebolavirus/fisiologia , Doença pelo Vírus Ebola/metabolismo , Infecções por Rickettsia/metabolismo , Rickettsia/fisiologia , Animais , Anexina A2/genética , Antígenos CD/genética , Antígenos CD/metabolismo , Caderinas/genética , Caderinas/metabolismo , Hemorragia Cerebral/genética , Hemorragia Cerebral/microbiologia , Hemorragia Cerebral/virologia , Endossomos/genética , Endossomos/metabolismo , Doença pelo Vírus Ebola/genética , Doença pelo Vírus Ebola/virologia , Humanos , Camundongos , Camundongos Knockout , Rickettsia/genética , Infecções por Rickettsia/genética , Infecções por Rickettsia/microbiologia
9.
J Cancer Res Clin Oncol ; 146(10): 2547-2557, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32671503

RESUMO

INTRODUCTION: Colorectal cancer (CRC) constitutes one of the most prevalent malignancies in the world. Recent research suggests that cancer stem cells (CSCs) are responsible for tumor cell's malignant behavior in CRC. This study has been designed to determinate clinical implications of CSC markers: CD44, DCLK1, Lgr5, and ANXA2 in CRC. MATERIALS AND METHODS: The study was performed on tissue samples which were collected from 89 patients undergoing colectomy. Formalin-fixed paraffin-embedded tissue blocks with representative tumor areas were identified and corded. Immunohistochemical staining was performed using anti-CD44, anti-LGR5, anti-ANXA2, and anti-DCLK1 antibodies. The H-score system was utilized to determine the immunointensity of CRC cells. RESULTS: The lower expression of Lgr5 was significantly correlated with the presence of lymph-node metastases (p = 0.011), while high expression of Lgr5 was statistically significant in vascular invasion in examined cancer tissue samples (p = 0.027). Moreover, a high H-score value of Lgr5 expression was significantly related to a reduced overall survival rate (p = 0.043). CONCLUSION: Our results suggest a strong relationship between CSC marker Lgr5 and vascular invasion, presence of lymph-node metastasis, and overall poor survival. The presence of Lgr5 might be an unfavorable prognostic factor, and its high level in cancer tissue is related to an aggressive course. This marker could also be used to access the effectiveness of the treatment.


Assuntos
Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Receptores Acoplados a Proteínas-G/metabolismo , Idoso , Anexina A2/metabolismo , Biomarcadores Tumorais/metabolismo , Neoplasias Colorretais/irrigação sanguínea , Progressão da Doença , Feminino , Humanos , Receptores de Hialuronatos/metabolismo , Imuno-Histoquímica , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Prognóstico , Proteínas Serina-Treonina Quinases/metabolismo , Receptores Acoplados a Proteínas-G/biossíntese , Análise Serial de Tecidos
10.
Life Sci ; 253: 117740, 2020 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-32376265

RESUMO

AIMS: Annexin A2 (ANXA2) is closely associated with tumor malignancy and its N-terminus includes a vital domain for its function. The aims are to explore the correlation between the sites (Tyr23, Ser1, Ser11 and Ser25) in the domain and its roles. MAIN METHODS: We re-expressed ANXA2 with mutated sites in ANXA2-deleted human colonic adenocarcinoma cell line caco2 (ANXA2-/-caco2). A series of analyses were used to determine the correlation of each site with ANXA2 activation, cell malignancy enhancement and motility-associated microstructural development. Bioinformatics and luciferase reporter assays were employed to validate ANXA2-targeted miRNAs. KEY FINDINGS: The in vitro results showed that all single and multiple mutations of the ANXA2 N-terminal sites inhibited ANXA2 phosphorylation at different levels and subsequently inhibited the proliferation, motility, and polymerization of F-actin and ß-tubulin in caco2 cells. Motility-associated microstructures were significantly remodeled when these sites were mutated. The forced expression of miR-206 significantly suppressed the proliferation, motility and epithelial-mesenchymal transition (EMT) of caco2 cells. The in vivo results showed that all the ANXA2 N-terminal site mutations and forced expression of miR-206 significantly inhibited tumor growth. Overall, this study demonstrated that the sites of the ANXA2 N-terminus, especially Tyr23, play crucial roles in maintaining the high malignancy of colonic adenocarcinoma. Furthermore, miR-206 targets ANXA2 and plays a role as a cancer suppressor in colonic adenocarcinoma. SIGNIFICANCE: Our study provided evidence that further elucidates the molecular mechanism of ANXA2 and its roles in colonic adenocarcinoma and suggested potential targets of ANXA2 for colonic adenocarcinoma therapy by using miR-206 as a novel strategy.


Assuntos
Adenocarcinoma/patologia , Anexina A2/genética , Neoplasias do Colo/patologia , MicroRNAs/genética , Actinas/metabolismo , Adenocarcinoma/genética , Animais , Células CACO-2 , Movimento Celular/genética , Proliferação de Células/genética , Neoplasias do Colo/genética , Transição Epitelial-Mesenquimal/genética , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Fosforilação/genética , Tubulina (Proteína)/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
11.
Nan Fang Yi Ke Da Xue Xue Bao ; 40(3): 382-387, 2020 Mar 30.
Artigo em Chinês | MEDLINE | ID: mdl-32376582

RESUMO

OBJECTIVE: To analyze the correlation of annexin A2 with coronary atherosclerotic heart disease (CAD) and the severity of CAD. METHODS: We collected data from a total of 200 inpatients admitted in our department between August, 2017 and August, 2019. According to the. RESULTS: of coronary angiography, the patients were divided into CAD group (n=150) and non-CAD (n=50), and the CAD patients was further divided, according to their clinical stability, into stable angina (SAP) group and acute coronary syndrome (ACS) group. Serum levels of annexin A2, MPO and PON1 were detected in all these patients, and their correlations with CAD, disease severity, and degree of coronary artery stenosis were analyzed.ResultsThe levels of annexin A2 and MPO were significantly higher in CAD patients than in non-CAD patients (P < 0.05). Among the CAD patients, those with ACS had significantly higher levels of annexin A2 (P < 0.05) and lower levels of PON-1 (P < 0.05) than those with SAP, but annexin A2 level was not significantly correlated with coronary lesion count, Gensini score, or the co-morbidity of diabetes. CONCLUSIONS: Annexin A2 is significantly elevated in patients with CAD, especially in those with ACS, and can be used as a predictor of clinical instability.


Assuntos
Anexina A2/sangue , Doença da Artéria Coronariana , Arildialquilfosfatase , Biomarcadores , Angiografia Coronária , Feminino , Humanos , Masculino , Índice de Gravidade de Doença
12.
Parasit Vectors ; 13(1): 183, 2020 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-32268913

RESUMO

BACKGROUND: Angiostrongylus cantonensis can cause severe symptoms of central nervous system infections. In the host, this parasite localizes in the blood and cerebrospinal fluid, and its secreted components can impact immune responses. Our previous study demonstrated that immune responses were inhibited in A. cantonensis-infected mice immunized with Ac-Galectin-1 (AcGal-1). However, the mechanisms by which AcGal-1 regulates the immune responses remain unclear. Macrophages are innate immune cells that rapidly respond to infection. The direct impact of AcGal-1 on macrophages may affect the immune responses. METHODS: AcGal-1 protein was purified by nickel ion affinity chromatography. The effect of AcGal-1 on the apoptosis of macrophages was detected using CCK-8 assay, flow cytometry and western blot. Macrophage membrane proteins bound to AcGal-1 were obtained using the His-tag-based pull-down assay and identified via mass spectrometry. Co-localization of AcGal-1 and the macrophage membrane protein Annexin A2 was observed by immunofluorescence microscopy, and their interaction was validated by co-immunoprecipitation experiments. SiRNA-mediated knockdown of Annexin A2 was used to determine if AcGal-1-induced macrophage apoptosis required interaction with Annexin A2. The phosphorylation level of apoptotic signal pathway protein was detected by phospho-antibody microarray and western blot. RESULTS: Our study showed that AcGal-1 caused apoptosis of the macrophages. AcGal-1 increased the expression of apoptosis proteins caspase-3, caspase-9, Bax, but reduced the expression of anti-apoptosis protein Bcl-2. AcGal-1 interacted with the membrane protein Annexin A2, and knockdown of Annexin A2 expression increased Bcl-2 but decreased Bax levels in AcGal-1-treated cells. Moreover, AcGal-1 increased JNK phosphorylation and the inhibition of JNK phosphorylation in AcGal-1-treated cells decreased the expression of caspase-3, -9, Bax and almost restored Bcl-2 to the level observed in control cells. CONCLUSIONS: AcGal-1 can induce the apoptosis of macrophages by binding to Annexin A2 and activating JNK downstream the apoptotic signaling pathway.


Assuntos
Anexina A2/imunologia , Apoptose , Galectina 1/imunologia , Sistema de Sinalização das MAP Quinases , Macrófagos/parasitologia , Angiostrongylus cantonensis , Animais , Proliferação de Células , Técnicas de Silenciamento de Genes , Humanos , Macrófagos/imunologia , Ligação Proteica , RNA Interferente Pequeno , Células THP-1
13.
J Vet Sci ; 21(2): e33, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32233139

RESUMO

Bovine ephemeral fever virus (BEFV) causes bovine ephemeral fever, which can produce considerable economic damage to the cattle industry. However, there is limited experimental evidence regarding the underlying mechanisms of BEFV. Annexin A2 (AnxA2) is a calcium and lipid-conjugated protein that binds phospholipids and the cytoskeleton in a Ca2+-dependent manner, and it participates in various cellular functions, including vesicular trafficking, organization of membrane domains, and virus proliferation. The role of the AnxA2 gene during virus infection has not yet been reported. In this study, we observed that AnxA2 gene expression was up-regulated in BHK-21 cells infected with the virus. Additionally, overexpression of the AnxA2 gene promoted the release of mature virus particles, whereas BEFV replication was remarkably inhibited after reducing AnxA2 gene expression by using the small interfering RNA (siRNA). For viral proteins, overexpression of the Matrix (M) gene promotes the release of mature virus particles. Moreover, the AnxA2 protein interaction with the M protein of BEFV was confirmed by GST pull-down and co-immunoprecipitation assays. Experimental results indicate that the C-terminal domain (268-334 aa) of AxnA2 contributes to this interaction. An additional mechanistic study showed that AnxA2 protein interacts with M protein and mediates the localization of the M protein at the plasma membrane. Furthermore, the absence of the AnxA2-V domain could attenuate the effect of AnxA2 on BEFV replication. These findings can contribute to elucidating the regulation of BEFV replication and may have implications for antiviral strategy development.


Assuntos
Anexina A2/metabolismo , Vírus da Febre Efêmera Bovina/fisiologia , Febre Efêmera/virologia , Proteínas da Matriz Viral/metabolismo , Replicação Viral , Animais , Bovinos , Regulação para Cima
14.
Nat Biomed Eng ; 4(3): 298-313, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32165732

RESUMO

The heterogeneity and continuous genetic adaptation of tumours complicate their detection and treatment via the targeting of genetic mutations. However, hallmarks of cancer such as aberrant protein phosphorylation and calcium-mediated cell signalling provide broadly conserved molecular targets. Here, we show that, for a range of solid tumours, a cyclic octapeptide labelled with a near-infrared dye selectively binds to phosphorylated Annexin A2 (pANXA2), with high affinity at high levels of calcium. Because of cancer-cell-induced pANXA2 expression in tumour-associated stromal cells, the octapeptide preferentially binds to the invasive edges of tumours and then traffics within macrophages to the tumour's necrotic core. As proof-of-concept applications, we used the octapeptide to detect tumour xenografts and metastatic lesions, and to perform fluorescence-guided surgical tumour resection, in mice. Our findings suggest that high levels of pANXA2 in association with elevated calcium are present in the microenvironment of most solid cancers. The octapeptide might be broadly useful for selective tumour imaging and for delivering drugs to the edges and to the core of solid tumours.


Assuntos
Anexina A2/metabolismo , Cálcio/metabolismo , Diagnóstico por Imagem/métodos , Neoplasias/diagnóstico por imagem , Células A549 , Animais , Anexina A2/genética , Apoptose , Linhagem Celular Tumoral , Modelos Animais de Doenças , Células HEK293 , Humanos , Macrófagos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Nus , Neoplasias/metabolismo , Neoplasias/patologia , Neoplasias Pancreáticas/diagnóstico por imagem , Fosforilação , Proteômica , Células Estromais , Transplante Heterólogo
15.
Fertil Steril ; 113(2): 364-373.e2, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32106990

RESUMO

OBJECTIVE: To demonstrate the feasibility of studying exosomes directly from peritoneal fluid, we isolated exosomes from endometriosis patient samples and from controls, and characterized their cargo. DESIGN: Case-control experimental study. SETTING: Academic clinical center. PATIENT (S): Women with and without endometriosis who underwent laparoscopic surgery (n = 28 in total). INTERVENTION (S): None. MAIN OUTCOME MEASURE (S): Concentration of exosomes within peritoneal fluid and protein content of the isolated exosomes. RESULT (S): Peritoneal fluid samples were pooled according to the cycle phase and disease stage to form six experimental groups, from which the exosomes were isolated. Exosomes were successfully isolated from peritoneal fluid in all the study groups. The concentration varied with cycle phase and disease stage. Proteomic analysis showed specific proteins in the exosomes derived from endometriosis patients that were absent in the controls. Five proteins were found exclusively in the endometriosis groups: PRDX1, H2A type 2-C, ANXA2, ITIH4, and the tubulin α-chain. CONCLUSION (S): Exosomes are present in peritoneal fluid. The characterization of endometriosis-specific exosomes opens up new avenues for the diagnosis and investigation of endometriosis.


Assuntos
Líquido Ascítico/química , Endometriose/metabolismo , Exossomos/química , Proteínas/análise , Adulto , Anexina A2/análise , Líquido Ascítico/patologia , Estudos de Casos e Controles , Endometriose/patologia , Exossomos/ultraestrutura , Estudos de Viabilidade , Feminino , Histonas/análise , Humanos , Pessoa de Meia-Idade , Peroxirredoxinas/análise , Proteínas Secretadas Inibidoras de Proteinases/análise , Proteômica , Tubulina (Proteína)/análise , Adulto Jovem
16.
Oncol Rep ; 43(2): 571-580, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31894281

RESUMO

Chemokine (C­C motif) ligand 18 (CCL18) is derived from breast tumor­associated macrophages (TAMs), which are primarily a macrophage subpopulation with an M2 phenotype. CCL18 binds to its receptor, PYK2 N­terminal domain interacting receptor 1 (Nir1), and promotes tumor progression and metastasis by inducing epithelial­mesenchymal transition (EMT) via the PI3K/Akt/GSK3ß/Snail signaling pathway in breast cancer cells. Recent research shows that Annexin A2 (AnxA2) plays a significant role in the invasion, metastasis, angiogenesis, proliferation, F­actin polymerization and multidrug resistance to chemotherapy of breast cancer. The present study aimed to elucidate the molecular mechanisms by which CCL18 promotes breast cancer progression through AnxA2 which are not fully understood. Western blot analysis showed that the expression of AnxA2 was upregulated in highly invasive breast cancer cell lines and invasive ductal carcinoma. Furthermore, through chemotaxis, scratch, Matrigel invasion, and spontaneous metastasis assays, it was demonstrated that AnxA2 enhanced the invasion of breast cancer cells and the metastasis of human breast cancer cells to lungs of SCID mice with CCL18 stimulation. Cellular F­actin measurement assay showed that reduction of AnxA2 suppressed CCL18­induced F­actin polymerization though phosphorylation of integrin ß1 in breast cancer cells. Immunofluorescence and western blot analysis revealed that AnxA2 promoted CCL18­induced EMT via the PI3K/Akt/GSK3ß/Snail signaling pathway, and LY294002 inhibited the phosphorylation of AnxA2 in vitro. In brief, AnxA2, as a downstream molecule of Nir 1 binding to CCL18, promotes invasion and metastasis by EMT through the PI3K/Akt/GSK3ß/Snail signaling pathway in breast cancer. This study suggests that AnxA2 is a potential anti­invasion/metastasis target for therapeutic intervention in breast cancer.


Assuntos
Anexina A2/metabolismo , Neoplasias da Mama/patologia , Carcinoma Ductal de Mama/patologia , Quimiocinas CC/metabolismo , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/secundário , Adulto , Idoso , Animais , Neoplasias da Mama/metabolismo , Carcinoma Ductal de Mama/metabolismo , Linhagem Celular Tumoral , Progressão da Doença , Transição Epitelial-Mesenquimal , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Pulmonares/metabolismo , Células MCF-7 , Camundongos , Pessoa de Meia-Idade , Transplante de Neoplasias , Carga Tumoral
17.
Cell Mol Life Sci ; 77(21): 4413-4428, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31894362

RESUMO

The chemotherapeutic drug epirubicin increases the exosomal export of miR-503 in endothelial cells. To understand the mechanisms behind this process, we transfected endothelial cells with miR-503 carrying a biotin tag. Then, we pulled-down the proteins interacting with miR-503 and studied their role in microRNA exosomal export. A total of four different binding partners were identified by mass spectrometry and validated by western blotting and negative controls, among them ANXA2 and hnRNPA2B1. Using knock-down systems combined with pull-down analysis, we determined that epirubicin mediates the export of miR-503 by disrupting the interaction between hnRNPA2B1 and miR-503. Then, both ANXA2 and miR-503 are sorted into exosomes while hnRNPA2B1 is relocated into the nucleus. The combination of these processes culminates in the increased export of miR-503. These results suggest, for the first time, that RNA-binding proteins can negatively regulate the exosomal sorting of microRNAs.


Assuntos
Células Endoteliais/metabolismo , Exossomos/metabolismo , Ribonucleoproteínas Nucleares Heterogêneas Grupo A-B/metabolismo , MicroRNAs/metabolismo , Anexina A2/metabolismo , Células Endoteliais da Veia Umbilical Humana , Humanos , Transporte de RNA
18.
Biochem Biophys Res Commun ; 523(3): 632-638, 2020 03 12.
Artigo em Inglês | MEDLINE | ID: mdl-31941608

RESUMO

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors, but its pathogenesis is not clear. This study found that the expression of TAGLN2 mRNA and protein in HCC was higher than that in adjacent tissues. TCGA database analysis further confirmed this result, and found that the expression of TAGLN2 was positively correlated with the prognosis of HCC, suggesting that TAGLN2 may be a tumor promoter gene. Then the TAGLN2-Annexin A2 (ANXA2) interaction and NF-κB signaling pathway were further clarified during the invasion and metastasis of HCC. This mechanism provides a theoretical basis for further finding molecular targets and drug targets related to HCC metastasis.


Assuntos
Anexina A2/genética , Carcinoma Hepatocelular/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/genética , Proteínas dos Microfilamentos/genética , Proteínas Musculares/genética , Anexina A2/análise , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Proliferação de Células , Progressão da Doença , Humanos , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/patologia , Proteínas dos Microfilamentos/análise , Pessoa de Meia-Idade , Proteínas Musculares/análise , Prognóstico , Regulação para Cima
19.
Breast Cancer Res ; 22(1): 11, 2020 01 28.
Artigo em Inglês | MEDLINE | ID: mdl-31992335

RESUMO

BACKGROUND: Limited information is available on biomarker(s) for triple-negative breast cancer (TNBC) that can address the higher incidence and aggressiveness of TNBC in African-American (AA) women. Our previous studies have demonstrated annexin A2 (AnxA2) association with exosomes which promotes angiogenesis and metastasis. Therefore, our goal was to examine the expression and function of exosomal-annexin A2 (exo-AnxA2) derived from the serum samples of breast cancer patients. METHODS: The expression of serum exo-AnxA2 and its association with clinicopathological features of the breast cancer patients were determined. The role of serum exo-AnxA2 to promote angiogenesis was determined by an in vivo Matrigel plug assay. RESULTS: Our results show that the expression of serum exo-AnxA2 in breast cancer patients (n = 169; 83.33 ± 2.040 ng/mL, P < 0.0001) is high compared to non-cancer females (n = 68; 34.21 ± 2.238 ng/mL). High expression of exo-AnxA2 levels in breast cancer was significantly associated with tumor grade (P < 0.0001), poor overall survival (hazard ratio (HR) 2.802; 95% confidence intervals (CI) = 1.030-7.620; P = 0.0353), and poor disease-free survival (HR 7.934; 95% CI = 1.778-35.398; P = 0.0301). The expression of serum exo-AnxA2 levels was significantly elevated in TNBC (n = 68; 109.1 ± 2.905 ng/mL; P < 0.0001) in comparison to ER+ (n = 50; 57.35 ± 1.545 ng/mL), HER2+ (n = 59; 78.25 ± 1.146 ng/mL), and non-cancer females (n = 68; 34.21 ± 2.238 ng/mL). Exo-AnxA2 showed diagnostic values with a maximum AUC as 1.000 for TNBC, 0.8304 for ER+, and 0.9958 for HER2+ compared to non-cancer females. The expression of serum exo-AnxA2 was significantly elevated in AA women with TNBC (n = 29; 118.9 ± 4.086 ng/mL, P < 0.0001) in comparison to Caucasian-American TNBC (n = 27; 97.60 ± 3.298 ng/mL) patients. Our in vivo results suggest a role of serum exo-AnxA2 in angiogenesis and its association with aggressiveness of TNBC in AA women. CONCLUSIONS: Our results demonstrated that the expression of serum exo-AnxA2 is high in AA women with TNBC and promotes angiogenesis. These findings suggest that exo-AnxA2 holds promise as a potential prognosticator of TNBC and may lead to an effective therapeutic option.


Assuntos
Afro-Americanos/estatística & dados numéricos , Anexina A2/sangue , Biomarcadores Tumorais/sangue , Exossomos/metabolismo , Neovascularização Patológica/patologia , Neoplasias de Mama Triplo Negativas/irrigação sanguínea , Animais , Feminino , Humanos , Camundongos , Camundongos Nus , Gradação de Tumores , Neovascularização Patológica/metabolismo , Curva ROC , Neoplasias de Mama Triplo Negativas/metabolismo , Neoplasias de Mama Triplo Negativas/patologia
20.
Int J Mol Sci ; 21(2)2020 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-31936170

RESUMO

Chimeric antigen receptors (CARs) have found clinical success in B cell malignancies, but a dearth of potential targets limits their wider clinical application, especially in solid tumours. Here, we describe the development of an anti-annexin A2 CAR, CAR(2448), derived from an antibody found to have activity against epithelial ovarian cancer cell lines. The spacer length of CAR(2448) was optimised based on in vitro cytotoxic activity against ovarian cancer (OC) cell lines via a real-time cytotoxicity assay. The longer spacer CAR(2448)L T cells exhibit significant effector activity, inducing inflammatory cytokine release and cytotoxicity against OC cell lines. Furthermore, CAR(2448)L-BBz T cells induced enhanced survival in an in vivo OC xenograft model and reduced tumour volume by 76.6%. Our preclinical studies of CAR(2448) suggest its potential for the unmet need of novel strategies for the treatment of ovarian cancer.


Assuntos
Anexina A2/imunologia , Carcinoma Epitelial do Ovário/terapia , Imunoterapia Adotiva , Neoplasias Ovarianas/terapia , Receptores de Antígenos Quiméricos/uso terapêutico , Animais , Anexina A2/antagonistas & inibidores , Carcinoma Epitelial do Ovário/imunologia , Linhagem Celular Tumoral , Feminino , Humanos , Imunoterapia Adotiva/métodos , Camundongos Endogâmicos NOD , Camundongos SCID , Neoplasias Ovarianas/imunologia , Receptores de Antígenos Quiméricos/imunologia , Linfócitos T/imunologia , Ensaios Antitumorais Modelo de Xenoenxerto
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