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1.
Int J Biol Sci ; 19(1): 258-280, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36594088

RESUMO

Background: Ovarian cancer (OC), a serious gynecological malignant disease, remains an enormous challenge in early diagnosis and medical treatment. Based on the GEO and TCGA databases in R language, endothelial cell-specific molecule 1 (ESM1) was confirmed separately with the bioinformatic analysis tool. ESM1 has been demonstrated to be upregulated in multiple cancer types, but the oncogenic mechanism by which ESM1 promotes OC is still largely unknown. Methods: In this study, we used WGCNA and random survival forest variable screening to filter out ESM1 in OC differentially expressed genes (DEGs). Next, we confirmed the mRNA and protein levels of ESM1 in OC samples via PCR and IHC. The correlation between the ESM1 level and clinical data of OC patients was further confirmed, including FIGO stage, lymph node metastasis, and recurrence. The role of ESM1 in OC development was explored by several functional experiments in vivo and in vitro. Then, the molecular mechanisms of ESM1 were further elucidated by bioinformatic end experimental analysis. Results: ESM1 was significantly upregulated in OC and was positively correlated with PFS but negatively correlated with OS. ESM1 knockdown inhibited cell proliferation, apoptosis escape, the cell cycle, angiogenesis, migration and invasion in multiple experiments. Moreover, GSVA found that ESM1 was associated with the Akt pathway, and our results supported this prediction. Conclusion: ESM1 was closely correlated with OC development and progression, and it could be considered a novel biomarker and therapeutic target for OC patients.


Assuntos
Neoplasias Ovarianas , Humanos , Feminino , Prognóstico , Neoplasias Ovarianas/metabolismo , Fatores de Transcrição , Metástase Linfática , Proteínas de Neoplasias , Proteoglicanas
2.
Int J Mol Sci ; 24(2)2023 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-36674659

RESUMO

Heparan sulfate is a ubiquitous, variably sulfated interactive glycosaminoglycan that consists of repeating disaccharides of glucuronic acid and glucosamine that are subject to a number of modifications (acetylation, de-acetylation, epimerization, sulfation). Variable heparan sulfate chain lengths and sequences within the heparan sulfate chains provide structural diversity generating interactive oligosaccharide binding motifs with a diverse range of extracellular ligands and cellular receptors providing instructional cues over cellular behaviour and tissue homeostasis through the regulation of essential physiological processes in development, health, and disease. heparan sulfate and heparan sulfate-PGs are integral components of the specialized glycocalyx surrounding cells. Heparan sulfate is the most heterogeneous glycosaminoglycan, in terms of its sequence and biosynthetic modifications making it a difficult molecule to fully characterize, multiple ligands also make an elucidation of heparan sulfate functional properties complicated. Spatio-temporal presentation of heparan sulfate sulfate groups is an important functional determinant in tissue development and in cellular control of wound healing and extracellular remodelling in pathological tissues. The regulatory properties of heparan sulfate are mediated via interactions with chemokines, chemokine receptors, growth factors and morphogens in cell proliferation, differentiation, development, tissue remodelling, wound healing, immune regulation, inflammation, and tumour development. A greater understanding of these HS interactive processes will improve therapeutic procedures and prognoses. Advances in glycosaminoglycan synthesis and sequencing, computational analytical carbohydrate algorithms and advanced software for the evaluation of molecular docking of heparan sulfate with its molecular partners are now available. These advanced analytic techniques and artificial intelligence offer predictive capability in the elucidation of heparan sulfate conformational effects on heparan sulfate-ligand interactions significantly aiding heparan sulfate therapeutics development.


Assuntos
Glicosaminoglicanos , Proteoglicanas , Glicosaminoglicanos/metabolismo , Proteoglicanas/metabolismo , Simulação de Acoplamento Molecular , Inteligência Artificial , Ligantes , Heparitina Sulfato/metabolismo
3.
BMC Gastroenterol ; 23(1): 4, 2023 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-36611136

RESUMO

BACKGROUND: Immune cells and stromal cells in the tumor microenvironment play a vital role in the progression of colorectal cancer (CRC). The study aimed to screen valuable prognostic biomarkers in CRC based on stromal and immune scores. METHOD: The ESTIMATE algorithm was used to calculate the immune and stromal scores of CRC samples in TCGA. Then samples were divided into high and low score groups based on the median value of the scores. Differentially expressed genes (DEGs) associated with immune and stromal scores were screened. WGCNA and univariate COX regression analysis were performed to further identify key prognostic genes. Analysis of scRNA-seq for CRC was used for verifying the main source of the key genes. The prognostic value of they was validated based on The Gene Expression Profiling Interactive Analysis and GSE17536 dataset. TIMER and CIBERSORT algorithms were applied to analyze the correlations among key genes and tumor-infiltrating immune cells. Several pairs of colon cancer tissue were used to be proven. RESULT: 1314 upregulated and 4 downregulated genes were identified, which were significantly enriched in immune-related biological processes and pathways. Among these DEGs, SPOCK1 and POSTN were identified as key prognostic genes and mainly expressed in cancer-associated fibroblasts for CRC. High expression of SPCOK1 and POSTN was associated with advanced clinical stage, T stage, N stage, and poor prognosis of CRC. The results from CIBERSORT and TIMER revealed that SPOCK1 and POSTN were associated with tumor-infiltrating immune cells, especially macrophages and neutrophils. Meanwhile, in several pairs of human colorectal tissue samples, SPOK1 and POSTN were found to be significantly overexpressed in colorectal tissue compared with para-cancer tissue, and macrophage surface markers CD68 (co-expressed by M1 and M2 macrophages) and CD206 (M2-specific macrophage expression) were also overexpressed in cancer tissue. Besides, SPOCK1 and POSTN expression were positively correlated with the expression of immune checkpoints. CONCLUSION: Collectively, our results indicate that SPOCK1 and POSTN associated with CAF may be novel prognostic biomarkers in CRC and correlate with immune infiltrates.


Assuntos
Neoplasias do Colo , Neoplasias Colorretais , Humanos , Prognóstico , Algoritmos , Perfilação da Expressão Gênica , Biomarcadores , Biomarcadores Tumorais/genética , Microambiente Tumoral/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Colorretais/genética , Moléculas de Adesão Celular/genética , Proteoglicanas
4.
Int J Biol Sci ; 19(2): 625-640, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36632458

RESUMO

Accumulating evidence shows that exosomes participate in cancer progression. However, the functions of cancer cell exosome-transmitted proteins are rarely studied. Previously, we reported that serglycin (SRGN) overexpression promotes invasion and metastasis of esophageal squamous cell carcinoma (ESCC) cells. Here, we investigated the paracrine effects of exosomes from SRGN-overexpressing ESCC cells (SRGN Exo) on ESCC cell invasion and tumor angiogenesis, and used mass spectrometry to identify exosomal proteins involved. Cation-dependent mannose-6-phosphate receptor (M6PR) and ephrin type-B receptor 4 (EphB4) were pronouncedly upregulated in SRGN Exo. Upregulated exosomal M6PR mediated the pro-angiogenic effects of SRGN Exo both in vitro and in vivo, while augmented exosomal EphB4 mediated the pro-invasive effect of SRGN Exo on ESCC cells in vitro. In addition, in vitro studies showed that manipulation of M6PR expression affected the viability and migration of ESCC cells. Both M6PR and EphB4 expression levels were positively correlated with that of SRGN in the serum of patients with ESCC. High level of serum M6PR was associated with poor overall survival rates. Taken together, this study presents the first proof that exosomal M6PR and EphB4 play essential roles in tumor angiogenesis and malignancy, and that serum M6PR is a novel prognostic marker for ESCC patients.


Assuntos
Neoplasias Esofágicas , Carcinoma de Células Escamosas do Esôfago , Exossomos , Humanos , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago/genética , Carcinoma de Células Escamosas do Esôfago/patologia , Exossomos/genética , Exossomos/metabolismo , Proteoglicanas/genética , Proteoglicanas/metabolismo , Neovascularização Patológica , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica
5.
Methods Mol Biol ; 2619: 3-24, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36662458

RESUMO

Proteoglycans (PGs) are macromolecules formed by a protein backbone to which one or more glycosaminoglycan (GAG) side chains are covalently attached. Most PGs are present in connective tissues, cell surfaces, and intracellular compartments. The major biological function of PGs derives from the GAG component of the molecule, which is involved in cell growth and proliferation, embryogenesis, maintenance of tissue hydration, and interactions of the cells via receptors. PGs are categorized into four groups based on their cellular and subcellular localization, including cell surfaces and extracellular, intracellular, and pericellular locations. GAGs are a crucial component of PGs involved in various physiological and pathological processes. GAGs also serve as biomarkers of metabolic diseases such as mucopolysaccharidoses and mucolipidoses. Detection of specific GAGs in various biological fluids helps manage various genetic metabolic disorders before it causes irreversible damage to the patient (Amendum et al., Diagnostics (Basel) 11(9):1563, 2021). There are several methods for detecting GAGs; this chapter focuses on measuring GAGs using enzyme-linked immunosorbent assay, liquid chromatographic tandem mass spectrometry, and automated high-throughput mass spectrometry.


Assuntos
Glicosaminoglicanos , Proteoglicanas , Humanos , Glicosaminoglicanos/química , Proteoglicanas/metabolismo , Cromatografia Líquida , Membrana Celular/metabolismo , Espectrometria de Massas em Tandem
6.
Methods Mol Biol ; 2619: 141-151, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36662468

RESUMO

Several experimental protocols are available to study the synthesis and secretion of proteoglycans in health and diseases, but there are few methods to analyse the intracellular processing of these macromolecules. We report a western blot analysis on medium and cell layer of primary chondrocyte culture to determine the glycanation status of aggrecan. Using a specific antibody against the aggrecan core protein and digesting an aliquot of sample with chondroitinase ABC, it is possible to analyse the whole aggrecan macromolecule and the core protein in order to evaluate defects in aggrecan glycanation.


Assuntos
Proteínas da Matriz Extracelular , Proteoglicanas , Agrecanas , Proteoglicanas/metabolismo , Técnicas de Cultura de Células , Western Blotting , Lectinas Tipo C
7.
Methods Mol Biol ; 2619: 99-106, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36662465

RESUMO

Glycosaminoglycans, the building blocks of proteoglycans, play a central role in the extracellular matrix and regulate a number of cellular processes. Therefore, any imbalance in their levels can lead to significant changes in cell behavior and phenotype. Additionally, glycosaminoglycans and their derivatives can be deployed as therapeutic agents in pathological conditions. Since cell morphology is a critical indicator of specialized cellular functions, its study can provide valuable insight. Scanning electron microscopy is a high-resolution imaging technique that makes for an ideal tool to observe the cellular appearance in 2D and 3D cultures under different conditions and/or substrates. In this chapter we provide a step-by-step protocol to study the influence of exogenously added glycosaminoglycans in the morphology of cells using scanning electron microscopy.


Assuntos
Glicosaminoglicanos , Proteoglicanas , Microscopia Eletrônica de Varredura , Matriz Extracelular/fisiologia
8.
Methods Mol Biol ; 2619: 125-139, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36662467

RESUMO

Extracellular vesicles (EVs) have emerged as a central mechanism of intercellular communication in physiology and disease. EVs participate in paracrine exchange of nucleic acids as well as lipids, proteins, and glycans to elicit a complex biological response in target cells. Proteoglycans (PGs) are widely expressed in EV-producing cells and are sorted to the membrane of secreted EVs to participate in some of the key processes in EV-mediated signaling. Most notably, PGs mainly of the heparan sulfate (HS) type are involved in EV biogenesis and cellular uptake of EVs through endocytic processes. EV-associated PGs may serve as short- and long-range chaperones of signaling molecules with potential implications for intercellular information exchange, most importantly in cancer development. This motivates the development of approaches targeting EV-HSPG interactions as a strategy in cancer treatment. Moreover, the importance of PG remodeling and alterations in their expression in cancer, together with the fact that EVs mimic their cell or tissue of origin, point at an important role of EV-associated PGs as disease biomarkers. Here, we provide methodological insights into the analysis of EV-PGs isolated from cell cultures as well as patient plasma liquid biopsy.


Assuntos
Vesículas Extracelulares , Neoplasias , Humanos , Proteoglicanas/metabolismo , Vesículas Extracelulares/metabolismo , Transdução de Sinais , Comunicação Celular , Neoplasias/patologia
9.
Methods Mol Biol ; 2619: 187-209, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36662471

RESUMO

Composite agarose-polyacrylamide gel electrophoresis (CAPAGE) in gels of 1.2% w/v polyacrylamide and 0.6% w/v agarose can be used to examine the heterogeneity of full-length native proteoglycan populations and their fragments in crude tissue extracts, and when used in conjunction with immunoblotting and specific antibodies to proteoglycan core protein and glycosaminoglycan, side chain epitopes can provide significant information on the level of proteoglycan polydispersity/heterogeneity and a number of proteoglycan populations present in tissue samples. This can be a technically difficult technique, but it reveals significant information on proteoglycans from small tissue samples not possible by any other separation methodology. Native full-length and proteoglycan fragments are examined in this technique something which cannot be done in the popular SDS-PAGE format unless the glycosaminoglycan side chains are first removed. Furthermore, since proteoglycans do not require renaturation from SDS-protein complexes, the proteoglycan populations separated by native electrophoresis are highly reactive with antibodies in immunoblotting procedures. Despite the massive sizes of proteoglycans, transfer conditions have been determined which provide close to quantitative transfer to nitrocellulose membranes without exceeding the binding capacity of such membranes, avoiding bleed-through of the transferred proteoglycans. Development of biotinylated hyaluronan and its application in an affinity blotting procedure has also yielded significant information on aggregatable proteoglycan populations separated by CAPAGE from a number of cartilages and vascular tissues in health and disease. While the CAPAGE system can be a technically demanding technique to master particularly in gel preparation, all other steps are straightforward, and the method yields invaluable information on proteoglycan populations extracted from connective tissues in health and disease that cannot be ascertained by any other technique. Further improvements in the detection of proteoglycan features with the development of novel bio-affinity probes or new antibody preparations are expected to further improve the utility of CAPAGE separation methodology.


Assuntos
Proteínas da Matriz Extracelular , Proteoglicanas , Proteoglicanas/metabolismo , Sefarose , Immunoblotting , Glicosaminoglicanos , Eletroforese em Gel de Poliacrilamida , Proteoglicanas de Sulfatos de Condroitina
10.
Methods Mol Biol ; 2619: 257-271, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36662476

RESUMO

Circulating microRNAs (miRNAs) possess significant roles in normal homeostasis and disease conditions, including cardiovascular diseases, fibrosis, inflammatory response, and cancer. Secreted miRNAs, via the membrane vesicles, actively communicate with extracellular matrix (ECM) components to affect cell-cell and cell-matrix interactions, thereby affecting matrix remodeling and metabolic pathways in the recipient cells. Matrix macromolecules regulate the expression of miRNAs, and in turn miRNAs have been identified as emerging mediators of matrix constituents, serving as appealing biomarkers for many pathophysiological processes. Therefore, the expression profile of certain miRNAs highlights the importance of their targeting in several aspects of human pathologies. In this chapter, we report molecular biology protocols to determine the effects of selected miRNAs on the expression and activity of matrix biomolecules.


Assuntos
MicroRNA Circulante , MicroRNAs , Neoplasias , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Proteoglicanas/metabolismo , Matriz Extracelular/metabolismo , Neoplasias/patologia
11.
Methods Mol Biol ; 2619: 273-292, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36662477

RESUMO

MicroRNAs are small noncoding RNAs that regulate gene expression at the posttranscriptional level. Proteoglycans are glycoproteins characterized by covalent attachment of a glycosaminoglycan chain, which have been identified as regulatory targets of microRNAs in a physiological and pathophysiological context. We present a strategy and detailed methods for the functional analysis of microRNA regulation of proteoglycans using human cancer cells as an application example. The experimental setup includes in silico microRNA target prediction, transfection of cancer cells with microRNA precursors, validation of target regulation by qPCR, flow cytometry and luciferase reporter assays, and an example for functional analysis and phenotype confirmation by complementation analysis.


Assuntos
MicroRNAs , Neoplasias , Humanos , MicroRNAs/metabolismo , Proteoglicanas/genética , Proteoglicanas/metabolismo , Transfecção , Luciferases/metabolismo
12.
Methods Mol Biol ; 2614: 261-271, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36587130

RESUMO

The extracellular matrix (ECM) is a molecular scaffold mainly comprising fibrous proteins, glycoproteins, proteoglycans, and polysaccharides. Aside from acting as a structural support, the ECM's composition dictates cell-matrix interactions at the biochemical and biophysical level. In the context of cancer, the ECM is a critical component of the tumor microenvironment (TME) and dysregulation of its deposition and remodelling has been shown to promote tumor onset, progression, and metastasis. Here, we describe a robust protocol for the isolation and subsequent proteomic analysis of the ECM of murine mammary glands, for downstream assays studying the role of the ECM in breast cancer. The protocol yields sufficient protein amounts to enable not only the global quantitation of protein expression but furthermore the enrichment and quantitative analysis of post-translationally modified peptides to study aberrant signalling.


Assuntos
Neoplasias da Mama , Glândulas Mamárias Humanas , Camundongos , Animais , Humanos , Feminino , Proteômica , Matriz Extracelular/metabolismo , Proteoglicanas/metabolismo , Neoplasias da Mama/patologia , Proteínas da Matriz Extracelular/metabolismo , Microambiente Tumoral
13.
Int J Biol Macromol ; 224: 1509-1523, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36550792

RESUMO

A proteoglycan LEPS1 was firstly isolated and purified from the spent substrate of Lentinula edodes, an agricultural waste that may cause environmental pollution. The average molecular weight of LEPS1 was 1.18 × 104 g/mol, and carbohydrate moiety (88.9 %) was composed of glucose, arabinose, galactose, xylose and mannose at a molar ratio of 1.2:1.2:1.0:2.3:1.1. The protein moiety (8.5 %) of LEPS1 was bonded to the polysaccharide chain via O-glycosidic linkage. LEPS1 could significantly improve the inflammatory injury of LPS stimulated RAW264.7 macrophages by inhibiting the secretion of NO and decreasing the levels of pro-inflammatory factors (TNF-α, IL-1ß and IL-6). LEPS1 inhibited JAK-STAT1 and p38 MAPK signaling pathway via modulating JAK expression, phosphorylation of STAT1 and phosphorylation of p38, respectively. Moreover, LEPS1 could promote the expression of CD 206 and IL-10 which were the markers for repairing macrophages. Overall, LEPS1 had anti-inflammatory activity and can potentially treat as a novel anti-inflammation agent. This work could provide scientific basis and valuable information for the highly efficient utilization of spent L. edodes substrates as the by-product in mushroom industries.


Assuntos
Cogumelos Shiitake , Cogumelos Shiitake/química , Proteoglicanas/metabolismo , Polissacarídeos/química , Anti-Inflamatórios/química , Macrófagos , Lipopolissacarídeos/farmacologia , Lipopolissacarídeos/metabolismo
14.
Methods Mol Biol ; 2598: 217-225, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36355295

RESUMO

Proteins from hyaline or articular cartilage can be isolated and purified using a series of chemical extraction steps and various identification techniques including mass spectrometry and immunoblotting. The isolation and purification of proteins from cartilage will facilitate the study of specific proteins and multimeric complexes of cartilage proteins to better understand their functions in normal healthy cartilage as well as pathological conditions of cartilage. Cartilage tissue engineering efforts rely on the comprehensive understanding of the composition of cartilage and the function of each of the protein constituents.


Assuntos
Cartilagem Articular , Cartilagem Hialina , Cartilagem Hialina/metabolismo , Cartilagem Articular/metabolismo , Proteoglicanas/metabolismo , Colágeno/metabolismo
15.
Biochem Biophys Res Commun ; 642: 185-191, 2023 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-36586186

RESUMO

Salmon nasal cartilage proteoglycan (PG) was orally administered to mice. The PG digest was recovered from the small intestine, and its sugar chain size and unsaturated disaccharide content were examined. The elution position of the PG digest following Sepharose CL-4B chromatography was consistent with that of actinase-digested PG prior to administration. The PG digest was incubated with chondroitinase ABC, which resulted in the elution pattern of the unsaturated disaccharides being identical to that of the degraded product of actinase-digested PG. The core protein of PG was digested in the mouse small intestine, but chondroitin sulfate, which is the sugar chain of PG, was not degraded at all. Then, the effects of chondroitin 4- and 6-sulfates on human colon cancer cells were examined. These chondroitin sulfates were found to suppress the expression of interleukin-6 induced by TNF-α. Overall, the chondroitin sulfate chain may act on the intestinal epithelium and suppress inflammation of the intestinal tract.


Assuntos
Sulfatos de Condroitina , Fator de Necrose Tumoral alfa , Camundongos , Humanos , Animais , Sulfatos de Condroitina/metabolismo , Interleucina-6 , Proteoglicanas/metabolismo , Condroitina , Dissacarídeos , Proteoglicanas de Sulfatos de Condroitina/metabolismo
16.
Methods Mol Biol ; 2557: 709-720, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36512246

RESUMO

Subcellular fractionation is an introductory step in a variety of experimental approaches designed to study intracellular components, like membranes and organelle systems. Subcellular fractions enriched in membranes of the Golgi apparatus of mammalian cells have been isolated to address localization and activity of proteins, including enzymes, to study intracellular membrane transport mechanisms, and to reconstitute in vitro cellular processes associated with the Golgi apparatus. Here, I describe methods to purify Golgi membranes by subcellular fractionation, to assay nucleotide sulfate (PAPS) uptake into Golgi vesicles, and to measure sulfate incorporation into in vitro synthesized glycosaminoglycans.


Assuntos
Fosfoadenosina Fosfossulfato , Proteoglicanas , Animais , Fosfoadenosina Fosfossulfato/metabolismo , Proteoglicanas/metabolismo , Complexo de Golgi/metabolismo , Glicosaminoglicanos/metabolismo , Sulfatos/metabolismo , Mamíferos/metabolismo
17.
PLoS One ; 17(12): e0267921, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36576921

RESUMO

Synovial fluid is composed of hyaluronan and proteoglycan-4 (PRG4 or lubricin), which work synergistically to maintain joint lubrication. In diseases like osteoarthritis, hyaluronan and PRG4 concentrations can be altered, resulting in lowered synovial fluid viscosity, and pro-inflammatory cytokine concentrations within the synovial fluid increase. Synovial fibroblasts within the synovium are responsible for contributing to synovial fluid and can be targeted to improve endogenous production of hyaluronan and PRG4 and to alter the cytokine profile. We cyclically loaded SW982 synoviocytes to 0%, 5%, 10%, or 20% strain for three hours at 1 Hz. To assess the impact of substrate stiffness, we compared the 0% strain group to cells grown on tissue culture plastic. We measured the expression of hyaluronan turnover genes, hyaluronan localization within the cell layer, hyaluronan concentration, PRG4 concentration, and the cytokine profile within the media. Our results show that the addition of cyclic loading increased HAS3 expression, but not in a magnitude-dependent response. Hyaluronidase expression was impacted by strain magnitude, which is exemplified by the decrease in hyaluronan concentration due to cyclic loading. We also show that PRG4 concentration is increased at 5% strain, while higher strain magnitude decreases overall PRG4 concentration. Finally, 10% and 20% strain show a distinct, more pro-inflammatory cytokine profile when compared to the unloaded group. Multivariate analysis showed distinct separation between certain strain groups in being able to predict strain group, hyaluronan concentration, and PRG4 concentration from gene expression or cytokine concentration data, highlighting the complexity of the system. Overall, this study shows that cyclic loading can be used tool to modulate the endogenous production of hyaluronan, PRG4, and cytokines from synovial fibroblasts.


Assuntos
Sinoviócitos , Sinoviócitos/metabolismo , Proteoglicanas/metabolismo , Ácido Hialurônico/metabolismo , Citocinas/metabolismo , Membrana Sinovial/metabolismo , Líquido Sinovial/metabolismo
18.
Nat Commun ; 13(1): 7704, 2022 12 13.
Artigo em Inglês | MEDLINE | ID: mdl-36513650

RESUMO

The epicardium, a mesothelial cell tissue that encompasses vertebrate hearts, supports heart regeneration after injury through paracrine effects and as a source of multipotent progenitors. However, the progenitor state in the adult epicardium has yet to be defined. Through single-cell RNA-sequencing of isolated epicardial cells from uninjured and regenerating adult zebrafish hearts, we define the epithelial and mesenchymal subsets of the epicardium. We further identify a transiently activated epicardial progenitor cell (aEPC) subpopulation marked by ptx3a and col12a1b expression. Upon cardiac injury, aEPCs emerge from the epithelial epicardium, migrate to enclose the wound, undergo epithelial-mesenchymal transition (EMT), and differentiate into mural cells and pdgfra+hapln1a+ mesenchymal epicardial cells. These EMT and differentiation processes are regulated by the Tgfß pathway. Conditional ablation of aEPCs blocks heart regeneration through reduced nrg1 expression and mesenchymal cell number. Our findings identify a transient progenitor population of the adult epicardium that is indispensable for heart regeneration and highlight it as a potential target for enhancing cardiac repair.


Assuntos
Traumatismos Cardíacos , Peixe-Zebra , Animais , Peixe-Zebra/metabolismo , Coração/fisiologia , Pericárdio , Células-Tronco/metabolismo , Traumatismos Cardíacos/genética , Transição Epitelial-Mesenquimal/genética , Proteoglicanas/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo
19.
Int J Mol Sci ; 23(24)2022 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-36555315

RESUMO

Glucocorticoids are steroid hormones that play diverse roles in numerous normal and pathological processes. They are actively used to treat a wide variety of diseases, including neurodegenerative and inflammatory diseases, cancers, and COVID-19, among others. However, the long-term use of glucocorticoids is associated with numerous side effects. Molecular mechanisms of these negative side effects are not completely understood. Recently, arguments have been made that one such mechanisms may be related to the influence of glucocorticoids on O-glycosylated components of the cell surface and extracellular matrix, in particular on proteoglycans and glycosaminoglycans. The potential toxic effects of glucocorticoids on these glycosylated macromolecules are particularly meaningful for brain physiology because proteoglycans/glycosaminoglycans are the main extracellular components of brain tissue. Here, we aim to review the known effects of glucocorticoids on proteoglycan expression and glycosaminoglycan content in different tissues, with a specific focus on the brain.


Assuntos
Glucocorticoides , Glicosaminoglicanos , Proteoglicanas , Humanos , Glucocorticoides/metabolismo , Glicosaminoglicanos/metabolismo , Proteoglicanas/metabolismo
20.
BMC Oral Health ; 22(1): 510, 2022 11 17.
Artigo em Inglês | MEDLINE | ID: mdl-36397112

RESUMO

BACKGROUND: Laryngeal cancer (LC) is the second frequent malignant head and neck cancer around world, while LC patients' prognosis is unsatisfactory. This study aims to investigate the prognostic value of tumor mutation burden (TMB)-related genes in LC. METHODS: LC data was downloaded from The Cancer Genome Atlas and Gene Expression Omnibus databases. TMB values of all samples were calculated basing on mutation data. The differentially expressed genes (DEGs) between LC samples with distinct TMB were subjected to univariate and LASSO Cox regression analysis to build Risk Score. Immune cell infiltration analysis was conducted in CIBERSORT. RESULTS: Between high and low TMB LC samples, we identified 210 DEGs. Of which, six optimal genes were included to construct Risk Score, comprising FOXJ1, EPO, FGF5, SPOCK1, KCNF1 and PSG5. High risk LC patients had significantly poorer overall survival than low risk patients. The nomogram model constructed basing on Risk Score and gender showed good performance in predicting LC patients' survival probability. CONCLUSIONS: The prognostic Risk Score model, basing on six TMB-related genes (FOXJ1, EPO, FGF5, SPOCK1, KCNF1 and PSG5), was a reliable prognostic model to separate LC patients with different prognoses.


Assuntos
Neoplasias Laríngeas , Humanos , Prognóstico , Neoplasias Laríngeas/genética , Regulação Neoplásica da Expressão Gênica , Biomarcadores Tumorais/genética , Fatores de Risco , Proteoglicanas
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