Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 5.777
Filtrar
1.
HNO ; 68(2): 100-105, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32006045

RESUMO

This manuscript describes the functional properties of the exosomes released from melanoma cells. It details the characteristics of the tumor antigen chondroitin sulfate proteoglycan 4 (CSPG4), which is used as a marker to separate exosomes released by melanoma cells from exosomes released by nonmalignant cells. The results are discussed in view of the potential role of melanoma cell-derived exosomes in the escape of malignant cells from the host's immune system.


Assuntos
Proteoglicanas de Sulfatos de Condroitina , Exossomos , Melanoma , Proteínas de Membrana , Neoplasias Cutâneas , Antígenos , Biomarcadores/análise , Líquidos Corporais , Proteoglicanas de Sulfatos de Condroitina/análise , Humanos , Melanoma/tratamento farmacológico , Melanoma/imunologia , Proteínas de Membrana/análise , Proteoglicanas , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/imunologia
2.
Zhonghua Gan Zang Bing Za Zhi ; 28(1): 83-86, 2020 Jan 20.
Artigo em Chinês | MEDLINE | ID: mdl-32023707

RESUMO

The early diagnosis and effective treatment of hepatocellular carcinoma (HCC) still remains a difficult problem that plagues the medical community. Exosomes are microvesicles with a diameter of 40~100 nm, and contains proteins, lipids and nucleic acids (mRNAs, lncRNAs, circRNAs, and microRNAs). They serve as an information exchange carrier, and play an important role in regulating and controlling the biomolecular function to maintain the stability of the intracellular environment. The function of exosomes in HCC includes intercellular communication, neoangiogenesis, cancer cell metastasis and multidrug resistance, which mediates the transformation of microRNAs (miRNA) and regulate the microenvironment of tumor progression, and then affect the pathophysiological behavior of cancer cells. Exosome-derived miRNA can be used for HCC monitoring or potential specific markers of early diagnosis. In addition, with the development and application prospects it could be a therapeutic goal for HCC. This paper summarizes the recent progress in the study of HCC-derived exosomal miRNA.


Assuntos
Carcinoma Hepatocelular/genética , Exossomos , Neoplasias Hepáticas/genética , MicroRNAs/genética , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , MicroRNAs/metabolismo , RNA Longo não Codificante , Microambiente Tumoral
3.
Anal Bioanal Chem ; 412(3): 601-609, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31897558

RESUMO

Numerous studies have shown that exosomes are closely related to the pathogenesis of various diseases, especially cancers. Therefore, a rapid and sensitive method for exosome detection will be of great importance for the diagnosis and prognosis of diseases. We report here a method for exosome detection based on the CD63 aptamer and clustered regular interspaced short palindromic repeats (CRISPR)/Cas12a system. This method consists mainly of exosomal membrane protein recognition based on the CD63 aptamer and signal amplification based on CRISPR/Cas12a. The CD63 aptamer, as an easily adaptable nucleic acid strand, is responsible for the conversion of the amounts of exosomes into nucleic acid detection, whereas CRISPR/Cas12a is responsible for highly specific nucleic acid signal amplification. The detection range of the method was determined as 3 × 103-6 × 107 particles per microliter. Additionally, we successfully applied this method to detect exosomes in clinical samples from both healthy individuals and patients with lung cancer, and the results were highly consistent with those obtained by nanoparticle tracking analysis. In general, this method provides a highly sensitive and specific method for the detection of exosomes and offers an avenue toward future exosome-based diagnosis of diseases.


Assuntos
Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Exossomos/química , Tetraspanina 30/análise , Células A549 , Sistemas CRISPR-Cas , Exossomos/patologia , Humanos , Neoplasias Pulmonares/patologia , Técnicas de Amplificação de Ácido Nucleico/métodos
4.
HNO ; 68(2): 106-110, 2020 Feb.
Artigo em Alemão | MEDLINE | ID: mdl-31915880

RESUMO

Due to late diagnosis and poor treatment response, advanced head and neck squamous cell carcinoma (HNSCC) belongs to the tumor diseases with highest mortality worldwide. Although many biomarkers have been investigated over the past years, none have yet become established in clinical practice. There is thus an urgent need to introduce noninvasive liquid biopsies that not only give information about cancer activity but also enable early conclusions regarding treatment response. This underlines the biological importance of exosomes from the blood of HNSCC patients. Isolation of exosomes from cell line supernatants and human plasma can easily be performed by size-exclusion chromatography. Thus, protein content, expression patterns, and immunomodulatory effects on immune cells can be evaluated. Further separation of exosomes by cell of origin enables more detailed examination of tumor-derived exosomes (TEX) and exosomes from immune cells. The etiology of the disease, e.g., human papillomavirus (HPV) status, disease activity (active vs. no evident disease), and response to immunotherapies can be detected by exosomal protein expression and immunosuppressive effects of exosomes on different immune cell subtypes. In conclusion, the presented studies can make an essential contribution to the establishment of exosomes as liquid biopsies for head and neck cancer diagnosis and treatment monitoring.


Assuntos
Exossomos , Neoplasias de Cabeça e Pescoço , Biópsia Líquida , Carcinoma de Células Escamosas de Cabeça e Pescoço , Neoplasias de Cabeça e Pescoço/diagnóstico , Humanos , Carcinoma de Células Escamosas de Cabeça e Pescoço/diagnóstico
5.
HNO ; 68(2): 71-79, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31965194

RESUMO

Exosomes are the current primary research focus of Dr. Theresa L. Whiteside. They are key mediators of intercellular communication in the head and neck, as well as other sites. Their effects in the tumor microenvironment are manifold and include suppression of immunity, promotion of angiogenesis, enabling of metastasis, as well as reprogramming of fibroblasts and mesenchymal stromal cells. The aim of this communication is to summarize Dr. Whiteside's contribution to the field of exosome research and details the interactions of exosomes with endothelial cells leading to recent findings on how to target endothelial cells using exosomes as a therapeutic approach.


Assuntos
Comunicação Celular , Exossomos , Neoplasias , Células Endoteliais , Humanos , Neovascularização Patológica , Microambiente Tumoral
6.
HNO ; 68(2): 111-114, 2020 Feb.
Artigo em Alemão | MEDLINE | ID: mdl-31996935

RESUMO

Until recently, exosomes were considered to be dust in electron microscopy. This perception has changed dramatically in the past few years. A new research field has emerged aiming to describe the formation, release, and function of these nanoparticles. The role of exosomes evolved from garbage bins to biologically active, selectively secreted particles that are part of a novel and broader messaging system. They were shown to display immunomodulatory properties. Today, exosomes are regarded as attractive targets for diagnostic and therapeutic purposes, especially in the field of oncology.


Assuntos
Exossomos , Nanopartículas , Microscopia Eletrônica
7.
Cancer Immunol Immunother ; 69(2): 285-292, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31897662

RESUMO

The wide-ranging collection of malignancies arising at the upper aerodigestive tract is categorized as head and neck cancer (HNC), the sixth most prevalent cancer worldwide. Infection with human papillomavirus (HPV) or exposure to carcinogens is the leading causes of HPV+ and HPV- HNCs development, respectively. HPV+ and HPV- HNCs are different in clinical and molecular aspects. Specifically, HPV- HNCs tightly associate with missense mutants of the TP53 gene (encoding for the p53 protein), suggesting a central role for mutant p53 gain-of-function (GOF) in driving tumorigenesis. In contrast, in HPV + HNC, the sequence of TP53 typically remains intact, while the protein is degraded. In tumor cells, the status of the TP53 gene affects the cargo of secreted exosomes. In this review, we describe the accumulated knowledge regarding the involvement of exosomes and p53 on cellular interactions between HPV+ and HPV- HNC cells, and the surrounding tumor microenvironment (TME). Moreover, we envision how TP53 status may determine exosomes cargo in HNC, and, consequently, modify the TME. The potential roles of exosomes described herein are based on both our studies and the studies of others on mutant p53-derived exosomes. Specifically, we showed how exosomes are shed by cancer cells harboring mutant p53 communicate with tumor-associated macrophages in the colon as well as with cancer-associated fibroblasts in the lung, creating immunosuppressive conditions and promoting invasiveness. Altogether, exosomes in HNC in the context of TP53 status are understudied and extensive research is required to shed light on the biology of HPV+ and HPV- HNC.


Assuntos
Transformação Celular Neoplásica/genética , Transformação Celular Neoplásica/metabolismo , Exossomos/metabolismo , Neoplasias de Cabeça e Pescoço/etiologia , Neoplasias de Cabeça e Pescoço/metabolismo , Mutação , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Animais , Suscetibilidade a Doenças , Matriz Extracelular/metabolismo , Predisposição Genética para Doença , Neoplasias de Cabeça e Pescoço/patologia , Humanos
8.
Life Sci ; 244: 117297, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31954745

RESUMO

As novel non-invasive tumor diagnostic biomarkers, exosomal bioactive miRNAs have received increasing attention. Herein, the aim of this study is to explore the clinical values and roles of exosomal miR106b in lung cancer. The exosomal miR-106b level was much higher in the serum of patients with lung cancer than that in healthy volunteers. Also, the exosomal miR-106b level in the lung cancer patient serum was associated with TNM stages and lymph node metastasis. Furthermore, exosomal miR-106b enhanced the migrated and invasive ability of lung cancer cells and increased the MMP-2 and MMP-9 expression. Mechanistically, exosomal miR-106b could target PTEN, and promote lung cancer cell migration and invasion. More importantly, PTEN overexpression reversed the effect of exosomal miR-106b on lung cancer cell migration and invasion. Taken together, these findings indicate that exosomal miR-106b may be a promising diagnostic biomarker and drug target for patients with lung cancer.


Assuntos
MicroRNAs/genética , PTEN Fosfo-Hidrolase/metabolismo , Adulto , Idoso , Apoptose/genética , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Exossomos/genética , Exossomos/metabolismo , Feminino , Expressão Gênica/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Metástase Linfática , Masculino , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Invasividade Neoplásica/genética , Metástase Neoplásica/genética , PTEN Fosfo-Hidrolase/genética , Prognóstico , Transdução de Sinais/genética
9.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 51(1): 81-86, 2020 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-31950794

RESUMO

Objective: To preliminarily investigate the differences of protein composition between immature dendritic cells (DC2.4) and their derived exosomes (DC-Exo) using a relatively rapid and sample-saving method based on nano-flow liquid chromatography tandem mass spectrometry (nanoLC-MS/MS). Methods: The supernatant of DC2.4 cells culture medium was collected and gradient centrifugation was applied to primarily extract and isolate DC-Exo; then sucrose density gradient ultracentrifugation was adopted to purify the DC-Exo. Bradford protein assay was used to determine the total protein content of the purified DC-Exo, and dynamic light scattering and transmission electron microscope were conducted to characterize the morphology and size distribution of the DC-Exo. Afterwards, protein samples including DC2.4 cells and DC-Exo were prepared by FASP enzymolysis method. Samples were performed nanoLC-MS/MS assay. The µLPickUp sample loading mode was used and only 1 µg of protein sample was required for each assay. The phase of Transport liquid and Micro A were both 0.05% trifluoroacetic acid-2% acetonitrile (ACN) aq. ( V/ V). Acclaim ® PepMap RSLC column was used to separate sample compositions and the gradient elute was adopted where the mobile phase consisted of (A) 0.1% formic acid (FA) and (B) 0.08% FA-80% ACN aq. ( V/ V) with flow rate of 0.3 µL/min. Positive APCI nanospray interface was used and "one-drive-ten" schema was set to collect primary information. The collected data was then searched and matched based on Uniport Mouse Fasta file as protein database in this case, and the re-annotated data was further sorted out and analyzed. Results: In the current study, relatively high yield of DC-Exo samples with sizes of 40-200 nm were obtained. The lyophilized protein samples prepared by FASP method could be loaded directly after redissolution, and only 1 µg of protein sample is required. The annotated results showed that DC2.4 cells contained 998 kinds of proteins, among which 227 were highly expressed and 535 were unique; while DC-Exo contained only 348 types of proteins, among which 18 were uniquely and highly expressed. There were 306 kinds of consensus proteins in both DC2.4 cells and DC-Exo, among them 7 kinds were highly expressed. Conclusion: The nanoLC-MS/MS method developed in this study only requires very small amount of protein samples, and it could primarily differentiate the protein compositions between DC2.4 cells and their derived exosomes rapidly.


Assuntos
Cromatografia Líquida , Células Dendríticas , Exossomos , Proteínas , Espectrometria de Massas em Tandem , Animais , Células Dendríticas/química , Exossomos/química , Camundongos , Proteínas/química
10.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 34(1): 124-131, 2020 Jan 15.
Artigo em Chinês | MEDLINE | ID: mdl-31939247

RESUMO

Objective: To investigate the effects of adipose-derived stem cell released exosomes (ADSC-Exos) on wound healing in diabetic mice. Methods: The ADSCs were isolated from the adipose tissue donated by the patients and cultured by enzymatic digestion. The supernatant of the 3rd generation ADSCs was used to extract Exos (ADSC-Exos). The morphology of ADSC-Exos was observed by transmission electron microscopy. The membrane-labeled proteins (Alix and CD63) were detected by Western blot, and the particle size distribution was detected by nanoparticle tracking analyzer. The fibroblasts were isolated from the skin tissue donated by the patients and cultured by enzymatic digestion. The 5th generation fibroblasts were cultured with PKH26-labeled ADSC-Exos, and observed by confocal fluorescence microscopy. The effects of ADSC-Exos on proliferation and migration of fibroblasts were observed with cell counting kit 8 (CCK-8) and scratch method. Twenty-four 8-week-old Balb/c male mice were used to prepare a diabetic model. A full-thickness skin defect of 8 mm in diameter was prepared on the back. And 0.2 mL of ADSC-Exos and PBS were injected into the dermis of the experimental group ( n=12) and the control group ( n=12), respectively. On the 1st, 4th, 7th, 11th, 16th, and 21st days, the wound healing was observed and the wound healing rate was calculated. On the 7th, 14th, and 21st days, the histology (HE and Masson) and CD31 immunohistochemical staining were performed to observe the wound structure, collagen fibers, and neovascularization. Results: ADSC-Exos were the membranous vesicles with clear edges and uniform size; the particle size was 40-200 nm with an average of 102.1 nm; the membrane-labeled proteins (Alix and CD63) were positive. The composite culture observation showed that ADSC-Exos could enter the fibroblasts and promote the proliferation and migration of fibroblasts. Animal experiments showed that the wound healing of the experimental group was significantly faster than that of the control group, and the wound healing rate was significantly different at each time point ( P<0.05). Compared with the control group, the wound healing of the experimental group was better. There were more microvessels in the early healing stage, and more deposited collagen fibers in the late healing stage. There were significant differences in the length of wound on the 7th, 14th, and 21st days, the number of microvessels on the 7th and 14th days, and the rate of deposited collagen fibers on the 14th and 21st days between the two groups ( P<0.05). Conclusion: ADSC-Exos can promote the wound healing in diabetic mice by promoting angiogenesis and proliferation and migration of fibroblasts and collagen synthesis.


Assuntos
Diabetes Mellitus Experimental , Exossomos , Adipócitos , Animais , Humanos , Masculino , Camundongos , Células-Tronco , Cicatrização
11.
Oral Dis ; 26(1): 173-181, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31630466

RESUMO

OBJECTIVES: Salivary exosomes harbour numerous constituents associated with oral and systemic diseases. However, no reports addressed components of salivary exosomes in patients with periodontitis. Our study aims to explore salivary exosomal proteins in young adults with severe periodontitis (SP) and to analyse the relationships between different proteins. MATERIALS AND METHODS: We collected saliva from 11 young adults with SP and 11 periodontally healthy subjects. After isolation of salivary exosomes, liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to analyse proteins. Gene ontology analysis was performed based on GeneCodis, and interaction network analysis for unique salivary exosomal proteins was performed by STRING. RESULTS: Twenty-six proteins were identified only in the SP group, and 58 proteins were identified only in the healthy group. Gene ontology analysis revealed that innate immune response, cytolysis and complement activation were highly enriched in the SP group. Interaction network analysis showed that the correlations among immune-related proteins (e.g. complement components and chemokine (C-C motif) ligand 28) were significant in the SP group. C6 proteins expressed only in the SP group were evaluated by Western blotting. CONCLUSIONS: Salivary exosomes from periodontitis patients are enriched immune-related proteins that might participate in the immune response during the development of periodontitis.


Assuntos
Exossomos/metabolismo , Periodontite/metabolismo , Proteínas e Peptídeos Salivares/metabolismo , Adulto , Estudos de Casos e Controles , Cromatografia Líquida , Humanos , Mapas de Interação de Proteínas , Saliva , Espectrometria de Massas em Tandem
12.
Oral Dis ; 26(1): 131-144, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31541596

RESUMO

OBJECTIVE: Secondary alveolar bone grafting is an essential part in the treatment of alveolar cleft deformity. Autologous iliac bone is the most favorable grafting source. However, the factors regulating postoperative bone formation are unclear. Investigations are needed to found whether the alveolar bone niche and bone marrow mesenchymal stem cells (BMSCs) derived from the jaw bone (BMSCs-J) affected the osteogenesis of BMSCs from the ilium (BMSCs-I). MATERIALS AND METHODS: The effect of BMSCs-J on BMSCs-I was investigated using a co-culture model. The exosomes were purified by sequential centrifugation. The osteoblastic differentiation of BMSCs was analyzed in vitro and in vivo. RESULTS: Co-culture with BMSCs-J increased the alkaline phosphatase (ALP) activity, Alizarin Red S (ARS) staining, and osteogenic gene expression in BMSCs-I. Transmission electron microscopy and nanoparticle tracking analysis verified the presence of exosomes in the culture supernatants of BMSCs. Exosomes secreted by BMSCs-J enhanced the ALP activity, ARS staining, osteogenic gene expression of BMSCs-I in vitro, and new bone formation in vivo. Blocking the secretion of exosomes using siRNA for Rab27a inhibited the effect of BMSCs-J. CONCLUSION: Exosomes played a role in the interaction between BMSCs-J and BMSCs-I, thereby leading to the enhanced osteogenic capacity of BMSCs-I and bone formation.


Assuntos
Células da Medula Óssea/citologia , Exossomos/fisiologia , Ílio/citologia , Células-Tronco Mesenquimais/citologia , Osteogênese , Fosfatase Alcalina/metabolismo , Diferenciação Celular , Células Cultivadas , Técnicas de Cocultura , Humanos , Arcada Osseodentária/citologia
13.
Cell Mol Life Sci ; 77(2): 253-265, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31468060

RESUMO

Dysregulation of angiogenesis is a phenomenon observed in several disorders such as diabetic foot, critical limb ischemia and myocardial infarction. Mesenchymal stromal cells (MSCs) possess angiogenic potential and have recently emerged as a powerful tool for cell therapy to promote angiogenesis. Although bone marrow-derived MSCs are the primary cell of choice, obtaining them has become a challenge. The placenta has become a popular alternative as it is a highly vascular organ, easily available and ethically more favorable with a rich supply of MSCs. Comparatively, placenta-derived MSCs (PMSCs) are clinically promising due to their proliferative, migratory, clonogenic and immunomodulatory properties. PMSCs release a plethora of cytokines and chemokines key to angiogenic signaling and facilitate the possibility of delivering PMSC-derived exosomes as a targeted therapy to promote angiogenesis. However, there still remains the challenge of heterogeneity in the isolated populations, questions on the maternal or fetal origin of these cells and the diversity in previously reported isolation and culture conditions. Nonetheless, the growing rate of clinical trials using PMSCs clearly indicates a shift in favor of PMSCs. The overall aim of the review is to highlight the importance of this rather poorly understood cell type and emphasize the need for further investigations into their angiogenic potential as an alternative source for therapeutic angiogenesis.


Assuntos
Células-Tronco Mesenquimais/fisiologia , Neovascularização Fisiológica/fisiologia , Placenta/fisiologia , Animais , Exossomos/fisiologia , Feminino , Humanos , Gravidez
15.
Recent Results Cancer Res ; 215: 319-344, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31605237

RESUMO

Extracellular micro- and nanoscale membrane vesicles produced by different cells progressively attract the attention of the scientific community. They function as mediators of intercellular communication and transport genetic material and signaling molecules between the cells. In the context of keeping homeostasis, the extracellular vesicles contribute to the regulation of various systemic and local processes. Vesicles released by the tumor and activated stromal cells exhibit multiple functions including support of tumor growth, preparation of the pre-metastatic niches, and immune suppression. Considerable progress has been made regarding the criteria of classification of the vesicles according to their origin, content, and function: Exosomes, microvesicles, also referred to as microparticles or ectosomes, and large oncosomes were defined as actively released vesicles. Additionally, apoptotic bodies represented by a highly heterogeneous population of particles produced during apoptosis, the programmed cell death, should be considered. Because the majority of isolation techniques do not allow the separation of different types of vesicles, a joined term "extracellular vesicles" (EVs) was recommended by the ISEV community for the definition of vesicles isolated from either the cell culture supernatants or the body fluids. Because EV content reflects the content of the cell of origin, multiple studies on EVs from body fluids in the context of cancer diagnosis, prediction, and prognosis were performed, actively supporting their high potential as a biomarker source. Here, we review the leading achievements in EV analysis from body fluids, defined as EV-based liquid biopsy, and provide an overview of the main EV constituents: EV surface proteins, intravesicular soluble proteins, EV RNA including mRNA and miRNA, and EV DNA as potential biomarkers. Furthermore, we discuss recent developments in technology for quantitative EV analysis in the clinical setting and future perspectives toward miniaturized high-precision liquid biopsy approaches.


Assuntos
Vesículas Extracelulares , Biópsia Líquida/métodos , Biópsia Líquida/tendências , Neoplasias/diagnóstico , Neoplasias/patologia , Apoptose , Micropartículas Derivadas de Células , Exossomos , Humanos
16.
Crit Rev Oncol Hematol ; 145: 102860, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31874447

RESUMO

Prostate cancer (PCa) is the most commonly diagnosed solid-organ cancer in males. The PSA testing may cause overdiagnosis and overtreatment for PCa patients. There is an urgent need for new biomarkers with greater discriminative precision for diagnosis and risk-stratification, to select for prostate biopsy and treatment of PCa. Liquid biopsy is a promising field with the potential to provide comprehensive information on the genetic landscape at diagnosis and to track genomic evolution over time in order to tailor the therapeutic choices at all stages of PCa. Exosomes, containing RNAs, DNAs and proteins, have been shown to be involved in tumour progression and a rich potential source of tumour biomarkers, especially for profiling analysis of their miRNAs content. In this review, we summarise the exosomal miRNAs in PCa diagnosis, prognosis and management, and further discuss their possible technical challenges associated with isolating PCa-specific exosomes.


Assuntos
Exossomos , Biópsia Líquida , MicroRNAs , Neoplasias da Próstata , Biomarcadores Tumorais , Biópsia , Humanos , Masculino , Neoplasias da Próstata/diagnóstico , Neoplasias da Próstata/genética
18.
J Enzyme Inhib Med Chem ; 35(1): 280-288, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31790614

RESUMO

Acidity, hypoxia and increased release of exosomes are severe phenotypes of tumours. The regulation of pH in tumours involves the interaction of several proteins, including the carbonic anhydrases which catalyze the formation of bicarbonate and protons from carbon dioxide and water. Among CA isoforms, CA IX is over-expressed in a large number of solid tumours, conferring to cancer cells a survival advantage in hypoxic and acidic microenvironment, but there isn't evidence that CA IX expression could have a real clinical impact. Therefore, in this study for the first time the expression and activity of CA IX have been investigated in the plasmatic exosomes obtained from patients with prostate carcinoma (PCa). For this purpose, the study was performed through different methodological approaches, such as NTA, western blot analysis, enzyme activity assay, Nanoscale flow cytometry, ELISA, confocal microscopy. The results showed that PCa exosomes significantly overexpressed CA IX levels and related activity as compared to healthy donors. Furthermore, CA IX expression and activity were correlated to the exosome intraluminal pH, demonstrating for the first time that PCa exosomes are acidic. Our data suggest the possible use of the exosomal CA IX expression and activity as a biomarker of cancer progression in PCa.


Assuntos
Antígenos de Neoplasias/biossíntese , Anidrase Carbônica IX/biossíntese , Exossomos/metabolismo , Neoplasias da Próstata/sangue , Idoso , Antígenos de Neoplasias/sangue , Anidrase Carbônica IX/sangue , Linhagem Celular , Humanos , Concentração de Íons de Hidrogênio , Masculino , Microscopia Confocal , Pessoa de Meia-Idade
19.
Biosci Biotechnol Biochem ; 84(1): 53-62, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31483222

RESUMO

Large numbers of miRNAs are found in biofluid exosomes. We isolated ~50-200 nm diameter exosomes from four types of porcine biofluid (urine, plasma, semen, and bile) using serial centrifugation and ultracentrifugation procedures. A total of 42.15 M raw data were generated from four small RNA libraries. This produced 40.17 M map-able sequences, of which we identified 204 conserved miRNAs, and 190 novel candidate miRNAs. Furthermore, we identified 34 miRNAs specifically expressed in only one library, all with well-characterized immune-related functions. A set of five universally abundant miRNAs (miR-148a-3p, miR-21-5p, let-7f-5p, let-7i-5p, and miR-99a-5p) across all four biofluids was also found. Function enrichment analysis revealed that the target genes of the five ubiquitous miRNAs are primarily involved in immune and RNA metabolic processes. In summary, our findings suggest that porcine biofluid exosomes contain a large number of miRNAs, many of which may be crucial regulators of the immune system.


Assuntos
Secreções Corporais , Líquidos Corporais , Exossomos/genética , Imunidade Inata , MicroRNAs/genética , MicroRNAs/imunologia , Suínos/genética , Animais , Sequência de Bases/genética , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/imunologia , Sequenciamento de Nucleotídeos em Larga Escala , Microscopia de Força Atômica , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de RNA
20.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 33(12): 1560-1565, 2019 Dec 15.
Artigo em Chinês | MEDLINE | ID: mdl-31823559

RESUMO

Objective: To investigate the effect of adipose-derived stem cell derived exosomes (ADSC-Exos) on angiogenesis after skin flap transplantation in rats. Methods: ADSCs were isolated and cultured by enzymatic digestion from voluntary donated adipose tissue of patients undergoing liposuction. The 3rd generation cells were observed under microscopy and identified by flow cytometry and oil red O staining at 14 days after induction of adipogenesis. After cells were identified as ADSCs, ADSC-Exos was extracted by density gradient centrifugation. And the morphology was observed by transmission electron microscopy, the surface marker proteins (CD63, TSG101) were detected by Western blot, and particle size distribution was measured by nanoparticle size tracking analyzer. Twenty male Sprague Dawley rats, weighing 250-300 g, were randomly divided into ADSC-Exos group and PBS group with 10 rats in each group. ADSC-Exos (ADSC-Exos group) and PBS (PBS group) were injected into the proximal, middle, and distal regions of the dorsal free flaps with an area of 9 cm×3 cm along the long axis in the two groups. The survival rate of the flap was measured on the 7th day, and then the flap tissue was harvested. The tissue morphology was observed by HE staining, and mean blood vessel density (MVD) was measured by CD31 immunohistochemical staining. Results: ADSCs were identified by microscopy, flow cytometry, and adipogenic induction culture. ADSC-Exos was a round or elliptical membrane vesicle with clear edge and uniform size. It has high expression of CD63 and TSG101, and its size distribution was 30-200 nm, which was in accordance with the size range of Exos. The distal necrosis of the flaps in the ADSC-Exos group was milder than that in the PBS group. On the 7th day, the survival rate of the flaps in the ADSC-Exos group was 64.2%±11.5%, which was significantly higher than that in the PBS group (31.0%±6.6%; t=7.945, P=0.000); the skin appendages in the middle region of the flap in the ADSC-Exos group were more complete, the edema in the proximal region was lighter and the vasodilation was more extensive. MVD of the ADSC-Exos group was (103.3±27.0) /field, which was significantly higher than that of the PBS group [(45.3±16.2)/field; t=3.190, P=0.011]. Conclusion: ADSC-Exos can improve the blood supply of skin flaps by promoting the formation of neovascularization after skin flap transplantation, thereby improve the survival rate of skin flaps in rats.


Assuntos
Tecido Adiposo , Exossomos , Transplante de Pele , Transplante de Células-Tronco , Adipócitos , Animais , Humanos , Masculino , Ratos , Ratos Sprague-Dawley , Células-Tronco , Retalhos Cirúrgicos
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA