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BACKGROUND: The family with sequence similarity 20-member C (FAM20C) kinase, a Golgi casein kinase, which is responsible for phosphorylating the majority of the extracellular phosphoproteins within S-x-E/pS motifs, and is fundamentally associated with multiple biological processes to maintain cell proliferation, biomineralization, migration, adhesion, and phosphate homeostasis. In dissecting how FAM20C regulates downstream molecules and potential mechanisms, however, there are multiple target molecules of FAM20C, particularly many phenomena remain elusive, such as changes in cell-autonomous behaviors, incompatibility in genotypes and phenotypes, and others. METHODS: Here, assay for transposase-accessible chromatin using sequencing (ATAC-seq), RNA sequencing (RNA-seq), proteomics, and phosphoproteomics were performed in Fam20c-dificient osteoblasts and to facilitate an integrated analysis and determine the impact of chromatin accessibility, genomic expression, protein alterations, signaling pathway, and post translational modifcations. RESULTS: By combining ATAC-seq and RNA-seq, we identified TCF4 and Wnt signaling pathway as the key regulators in Fam20c-dificient cells. Further, we showed Calpastatin/Calpain proteolysis system as a novel target axis for FAM20C to regulate cell migration and F-actin cytoskeleton by integrated analysis of proteomics and phosphoproteomics. Furthermore, Calpastatin/Calpain proteolysis system could negatively regulate the Wnt signaling pathway. CONCLUSION: These observations implied that Fam20c knockout osteoblasts would cause cell homeostatic imbalance, involving changes in multiple signaling pathways in the conduction system.
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Calpaína , Proteínas da Matriz Extracelular , Proteínas da Matriz Extracelular/genética , Proteólise , Calpaína/metabolismo , Movimento Celular , HomeostaseRESUMO
Crumb rubber (CR) is a commonly used infill material in artificial turf worldwide. However, the potential health risk associated with exposure to CR containing environmentally persistent free radicals (EPFRs) remains under investigation. Herein, we observed the widespread presence of CR particles in the range of 2.8-51.4 µg/m3 and EPFRs exceeding 6 × 1015 spins/g in the ambient air surrounding artificial turf fields. Notably, the abundance of these particles tended to increase with the number of operating years of the playing fields. Furthermore, by analyzing saliva samples from 200 participants, we established for the first time that EPFR-carrying CR could be found in saliva specimens, suggesting the potential for inhaling them through the oral cavity and their exposure to the human body. After 40 min of exercise on the turf, we detected a substantial presence of EPFRs, reaching as high as (1.15 ± 1.00) × 1016 spins of EPFR per 10 mL of saliva. Moreover, the presence of EPFRs considerably increased the oxidative potential of CR, leading to the inactivation of Ca2+, redox reactions, and changes in spatial binding of the α-1,4-chain of salivary amylase to Ca2+, all of which could influence human saliva health. Our study provides insights into a new pathway of human exposure to CR with EPFRs in artificial turf infill, indicating an increased human health risk of CR exposure.
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Exposição Ambiental , Borracha , Humanos , Exposição Ambiental/análise , Saliva , Radicais LivresRESUMO
Background: Lung ischemia-reperfusion injury (LIRI) remains the major cause of primary lung dysfunction after lung transplantation. Diabetes mellitus (DM) is an independent risk factor for morbidity and mortality following lung transplantation. Mitochondrial dysfunction is recognized as a key mediator in the pathogenesis of diabetic LIRI. Melatonin has been reported to be a safe and potent preserving mitochondrial function agent. This study aimed at investigating the potential therapeutic effect and mechanisms of melatonin on diabetic LIRI. Methods: High-fat-diet-fed streptozotocin-induced type 2 diabetic rats were exposed to melatonin, with or without administration of the SIRT3 short hairpin ribonucleic acid (shRNA) plasmid following a surgical model of ischemia-reperfusion injury of the lung. Lung function, inflammation, oxidative stress, cell apoptosis, and mitochondrial function were examined. Results: The SIRT3 signaling and mitophagy were suppressed following diabetic LIRI. Treatment with melatonin markedly induced mitophagy and restored SIRT3 expression. Melatonin treatment also attenuated subsequent diabetic LIRI by improving lung functional recovery, suppressing inflammation, decreasing oxidative damage, diminishing cell apoptosis, and preserving mitochondrial function. However, either administration of SIRT3 shRNA or an autophagy antagonist 3-methyladenine (3-MA) suppressing mitophagy, and compromised the protective action of melatonin. Conclusion: Data indicated that melatonin attenuates diabetic LIRI through activation of SIRT3 signaling-mediated mitophagy.
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Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 2 , Melatonina , Traumatismo por Reperfusão , Sirtuína 3 , Ratos , Animais , Sirtuína 3/metabolismo , Sirtuína 3/farmacologia , Sirtuína 3/uso terapêutico , Melatonina/farmacologia , Melatonina/uso terapêutico , Melatonina/metabolismo , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/patologia , Mitofagia , Traumatismo por Reperfusão/tratamento farmacológico , Pulmão/metabolismo , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/tratamento farmacológico , RNA Interferente Pequeno/metabolismo , ApoptoseRESUMO
OBJECTIVE: Histone-deacetylases (HDACs) are epigenetic modulators involved in the control of gene expression. No data are available on the expression or subcellular localization of HDACs in salivary glands. The present study aims to examine the subcellular distribution of HDACs in salivary glands during postnatal development. DESIGN: The major salivary glands of C57/BL6 mice were separately removed at 10, 25, 30,60 and 90 days after birth. Hematoxylin-eosin (H&E) and immunohistochemical staining were performed for HDACs. Gene Expression of HDACs in C57BL/6. NOD-Aec1Aec2 mice salivary glands during the development of Sjögren's syndrome-like illness were also analyzed by using the gene expression datasets (GSE 15640). RESULTS: In the mice salivary gland, HDACs were found to have different localization patterns at various stages of development (10, 25, 30, 60, and 90 days). Apart from HDAC6, ductal cells of salivary glands were the primary sites for HDAC localization. HDAC2, 8, 5, 10 and 11 were expressed at high levels in the salivary gland after birth while HDAC6 showed no expression during postnatal development. This suggests that these HDAC subtypes may have different roles in salivary gland function. In the context of Sjögren's syndrome-like illness, HDAC 2, 8 and 10 showed low expression while HDAC1, 6,5,3 and 11 had relatively high expression in the salivary gland. CONCLUSIONS: This study has provided an important reference for understanding the spatiotemporal-specific expression of HDACs in the salivary gland. These results offer new clues for the experimenters and hold promise for developing innovative therapeutic strategies for salivary gland-related diseases.
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Síndrome de Sjogren , Animais , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos NOD , Histona Desacetilases/genética , Histonas , Glândulas SalivaresRESUMO
BACKGROUND: Acute lung injury (ALI) is a leading cause of mortality in patients with sepsis due to proinflammatory endothelial changes and endothelial permeability defects. Mitochondrial dysfunction is recognized as a critical mediator in the pathogenesis of sepsis-induced ALI. Although mitophagy regulation of mitochondrial quality is well recognized, little is known about its role in lung ECs during sepsis-induced ALI. Sirtuin 1 (SIRT1) is a histone protein deacetylase involved in inflammation, mitophagy, and cellular senescence. Here, the authors show a type of late endosome-dependent mitophagy that inhibits NLRP3 and STING activation through SIRT1 signaling during sepsis-induced ALI. METHODS: C57BL/6J male mice with or without administration of the SIRT1 inhibitor EX527 in the CLP model and lung ECs in vitro were developed to identify mitophagy mechanisms that underlie the cross-talk between SIRT1 signaling and sepsis-induced ALI. RESULTS: SIRT1 deficient mice exhibited exacerbated sepsis-induced ALI. Knockdown of SIRT1 interfered with mitophagy through late endosome Rab7, leading to the accumulation of damaged mitochondria and inducing excessive mitochondrial reactive oxygen species (mtROS) generation and cytosolic release of mitochondrial DNA (mtDNA), which triggered NLRP3 inflammasome and the cytosolic nucleotide sensing pathways (STING) over-activation. Pharmacological inhibition of STING and NLRP3 i n vivo or genetic knockdown in vitro reversed SIRT1 deficiency mediated endothelial permeability defects and endothelial inflammation in sepsis-induced ALI. Moreover, activation of SIRT1 with SRT1720 in vivo or overexpression of SIRT1 in vitro protected against sepsis-induced ALI. CONCLUSION: These findings suggest that SIRT1 signaling is essential for restricting STING and NLRP3 hyperactivation by promoting endosomal-mediated mitophagy in lung ECs, providing potential therapeutic targets for treating sepsis-induced ALI.
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Lesão Pulmonar Aguda , Proteínas de Membrana , Camundongos Endogâmicos C57BL , Mitofagia , Proteína 3 que Contém Domínio de Pirina da Família NLR , Sepse , Sirtuína 1 , Animais , Sirtuína 1/metabolismo , Sirtuína 1/antagonistas & inibidores , Lesão Pulmonar Aguda/metabolismo , Lesão Pulmonar Aguda/etiologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/antagonistas & inibidores , Sepse/metabolismo , Sepse/complicações , Mitofagia/fisiologia , Masculino , Camundongos , Proteínas de Membrana/metabolismo , Proteínas rab de Ligação ao GTP/metabolismo , Transdução de Sinais/fisiologia , Endossomos/metabolismo , Modelos Animais de DoençasRESUMO
Family with sequence similarity 20 member C (FAM20C) is a Golgi casein kinase that phosphorylates extracellularly-secreted regulatory proteins involved in bone development and mineralization, but its specific role in bone development is still largely unknown. In this study, to examine the specific mechanisms that FAM20C influences bone development, we cross-bred Osx-Cre with FAM20Cflox/flox mice to establish a Osx-Cre; FAM20Cflox/flox knockout (oKO) mouse model; FAM20C was KO in pre-osteoblasts. oKO development was examined at 1-10 weeks, in which compared to control FAM20Cflox/flox, they had lower body weights and bone tissue mineralization. Furthermore, oKO had lower bone volume fractions, thickness, and trabecular numbers, along with higher degrees of trabecular separation. These mice also had decreased femoral metaphyseal cartilage proliferation layer, along with thickened hypertrophic layer and increased apoptotic cell counts. Transcriptomic analysis found that differentially-expressed genes in oKO were concentrated in the osteoclast differentiation pathway, in line with increased osteoclast presence. Additionally, up-regulation of osteoclast-related, and down-regulation of osteogenesis-related genes, were identified, in which the most up-regulated genes were signal regulatory protein ß-1 family (Sirpb1a-c) and mitogen-activated protein kinase 13. Overall, FAM20C KO in pre-osteoblasts leads to abnormal long bone development, likely due to subsequent up-regulation of osteoclast differentiation-associated genes.
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Desenvolvimento Ósseo , Proteínas de Ligação ao Cálcio , Caseína Quinase I , Diferenciação Celular , Camundongos Knockout , Osteoblastos , Osteoclastos , Osteogênese , Regulação para Cima , Animais , Camundongos , Desenvolvimento Ósseo/genética , Caseína Quinase I/metabolismo , Caseína Quinase I/genética , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Osteoblastos/metabolismo , Osteoclastos/metabolismo , Osteogênese/genética , Masculino , FemininoRESUMO
Hydrogels are widely used as wound dressings for wound healing, but when hydrogels absorb wound exudate, swelling occurs and compresses the surrounding tissue, affecting healing. A chitosan injectable (CS/4-PA/CAT) hydrogel based on catechol and 4-glutenoic acid was prepared to avoid swelling and promote wound healing. After cross-linking by UV light, pentenyl groups formed hydrophobic alkyl chains which give the hydrogel a hydrophobic network and thus control its swelling. CS/4-PA/CAT hydrogels retained their non-swelling for a long time in PBS solution at 37 °C. CS/4-PA/CAT hydrogels had good injectable and adhesive properties, and had a good killing effect on E. coli and S. aureus and could remove the bacterial biofilms of E. coli and S. aureus. CS/4-PA/CAT hydrogels had good in vitro coagulation function by absorbing red blood cells and platelets. When used in a whole skin injury model, CS/4-PA/CAT-1 hydrogel stimulated fibroblast migration, promoted epithelialization and accelerated collagen deposition to promote defect healing, and showed good hemostatic effects in liver and femoral artery defects in mice. In summary, the non-swelling injectable hydrogel with free radical scavenging, rapid hemostasis, and antibacterial effects would be a promising treatment for defect repair.
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Quitosana , Camundongos , Animais , Quitosana/química , Cicatrização/fisiologia , Raios Ultravioleta , Hidrogéis/farmacologia , Hidrogéis/química , Staphylococcus aureus , Escherichia coli , Antibacterianos/farmacologia , Antibacterianos/químicaRESUMO
BACKGROUND: Cell survival or death is one of the key scientific issues of inflammatory response. To regulate cell death during the occurrence and development of periodontitis, various forms of programmed cell death, such as pyroptosis, ferroptosis, necroptosis, and apoptosis, have been proposed. It has been found that ferroptosis characterized by iron-dependent lipid peroxidation is involved in cancer, degenerative brain diseases and inflammatory diseases. Furthermore, NCOA4 is considered one of ferroptosis-related genes (FRGs) contributing to butyrate-induced cell death in the periodontitis. This research aims to analyze the expression of FRGs in periodontitis tissues and to explore the relationship between ferroptosis and periodontitis. METHOD: Genes associated with periodontitis were retrieved from two Gene Expression Omnibus datasets. Then, we normalized microarray data and removed the batch effect using the R software. We used R to convert the mRNA expression data and collected the expression of FRGs. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), transcription factor (TF) and protein-protein interaction (PPI) network analyses were used. In addition, we constructed a receiver operating characteristic curve and obtained relative mRNA expression verified by quantitative reverse-transcription polymerase chain reaction (PCR). RESULTS: Eight and 10 FRGs related to periodontitis were upregulated and downregulated, respectively. GO analysis showed that FRGs were enriched in the regulation of glutathione biosynthetic, glutamate homeostasis, and endoplasmic reticulum-nucleus signaling pathway. The top TFs included CEBPB, JUND, ATF2. Based on the PPI network analysis, FRGs were mainly linked to the negative regulation of IRE1-mediated unfolded protein response, regulation of type IIa hypersensitivity, and regulation of apoptotic cell clearance. The expression levels of NCOA4, SLC1A5 and HSPB1 using PCR were significantly different between normal gingival samples and periodontitis samples. Furthermore, the diagnostic value of FRGs for periodontitis were "Good". CONCLUSIONS: We found significant associations between FRGs and periodontitis. The present study not only provides a new possible pathomechanism for the occurrence of periodontitis but also offers a new direction for the diagnosis and treatment of periodontitis.
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Ferroptose , Periodontite , Sistema ASC de Transporte de Aminoácidos , Biologia Computacional , Ferroptose/genética , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Humanos , Antígenos de Histocompatibilidade Menor , Periodontite/genética , Periodontite/metabolismo , RNA Mensageiro/genéticaRESUMO
INTRODUCTION: Aspirin is a common antipyretic, analgesic, and anti-inflammatory drug, which has been reported to extend life in animal models and application in the treatment of aging-related diseases. However, it remains unclear about the effects of aspirin on bone marrow-derived mesenchymal stromal cells (BM-MSCs). Here, we aimed to analyze the influence of aspirin on senescence and young BM-MSCs. METHODS: BM-MSCs were serially passaged to construct a replicative senescence model. SA-ß-gal staining, PCR, western blot, and RNA-sequencing were performed on BM-MSCs with or without aspirin treatment, to examine aspirin's impact on bone marrow-derived mesenchymal stem cells. RESULTS: SA-ß-gal staining, PCR, and western blot revealed that aspirin could alleviate the cellular expression of senescence-related indicators of BM-MSCs, including a decrease of SA-ß-gal-positive cells and staining intensity, and downregulation of p16, p21, and p53 expression after aspirin treatment. RNA-sequencing results shown in the biological processes related to aging, aspirin could influence cellular immune response and lipid metabolism. CONCLUSION: The efficacy of aspirin for retarding senescence of BM-MSCs was demonstrated. Our study indicated that the mechanisms of this delay might involve influencing immune response and lipid metabolism.
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Senescência Celular , Células-Tronco Mesenquimais , Animais , Senescência Celular/genética , Aspirina/farmacologia , Medula Óssea , RNA-Seq , Células Cultivadas , RNA/metabolismoRESUMO
BACKGROUND: The family with sequence similarity 20-member C (Fam20C) kinase plays important roles in physiopathological process and is responsible for majority of the secreted phosphoproteome, including substrates associated with tumor cell migration. However, it remains unclear whether Fam20C plays a role in cancers. Here, we aimed to analyze the expression and prognostic value of Fam20C in pan-cancer and to gain insights into the association between Fam20C and immune infiltration. METHODS: We analyzed Fam20C expression patterns and the associations between Fam20C expression levels and prognosis in pan-cancer via the ONCOMINE, TIMER (Tumor Immune Estimation Resource), PrognoScan, GEPIA (Gene Expression Profiling Interactive Analysis), and Kaplan-Meier Plotter databases. After that, GEPIA and TIMER databases were applied to investigate the relations between Fam20C expression and immune infiltration across different cancer types, especially BLCA (bladder urothelial carcinoma), LGG (brain lower grade glioma), and STAD (stomach adenocarcinoma). RESULTS: Compared with adjacent normal tissues, Fam20C was widely expressed across many cancers. In general, Fam20C showed a detrimental role in pan-cancer, it was positively associated with poor survival of BLCA, LGG, and STAD patients. Specifically, based on TCGA (The Cancer Genome Atlas) database, a high expression level of Fam20C was associated with worse prognostic value in stages T2-T4 and stages N0-N2 in the cohort of STAD patients. Moreover, Fam20C expression had positive associations with immune infiltration, including CD4+ T cells, macrophages, neutrophils, and dendritic cells, and other diverse immune cells in BLCA, LGG, and STAD. CONCLUSION: Fam20C may serve as a promising prognostic biomarker in pan-cancer and has positive associations with immune infiltrates.
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Caseína Quinase I/fisiologia , Proteínas da Matriz Extracelular/fisiologia , Neoplasias/imunologia , Caseína Quinase I/genética , Proteínas da Matriz Extracelular/genética , Humanos , Estimativa de Kaplan-Meier , Metástase Linfática , Neoplasias/genética , Neoplasias/patologia , Oncogenes , Prognóstico , RNA Mensageiro/genéticaRESUMO
BACKGROUND: Neurogenic heterotopic ossification is a disorder of aberrant bone formation affecting one in five patients sustaining a spinal cord injury or traumatic brain injury (SCI-TBI-HO). However, the underlying mechanisms of SCI-TBI-HO have proven difficult to elucidate. The aim of the present study is to identify the most promising candidate genes and biological pathways for SCI-TBI-HO. METHODS: In this study, we used text mining to generate potential explanations for SCI-TBI-HO. Moreover, we employed several additional datasets, including gene expression profile data, drug data and tissue-specific gene expression data, to explore promising genes that associated with SCI-TBI-HO. RESULTS: We identified four SCI-TBI-HO-associated genes, including GDF15, LDLR, CCL2, and CLU. Finally, using enrichment analysis, we identified several pathways, including integrin signaling, insulin pathway, internalization of ErbB1, urokinase-type plasminogen activator and uPAR-mediated signaling, PDGFR-beta signaling pathway, EGF receptor (ErbB1) signaling pathway, and class I PI3K signaling events, which may be associated with SCI-TBI-HO. CONCLUSIONS: These results enhance our understanding of the molecular mechanisms of SCI-TBI-HO and offer new leads for researchers and innovative therapeutic strategies.
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Family with sequence similarity 20member C (FAM20C), a recently characterized Golgi kinase, performs numerous biological functions by phosphorylating more than 100 secreted proteins. However, the role of FAM20C in the salivary glands remains undefined. The present study demonstrated that FAM20C is mainly located in the cytoplasm of duct epithelial cells in the salivary glands. Fam20cf/f; MmtvCre mice were created in which Fam20c was inactivated in the salivary gland cells and observed that the number of ducts and the ductal crosssectional area increased significantly, while the number of acinar cells was reduced. The granular convoluted tubules (GCTs) exhibited an accumulation of aberrant secretory granules, along with a reduced expression and altered distribution patterns of ß nerve growth factor, αamylase and bone morphogenetic protein (BMP) 4. This abnormality suggested that the GCT cells were immature and exhibited defects in developmental and secretory functions. In accordance with the morphological alterations and the reduced number of acinar cells, FAM20C deficiency in the salivary glands significantly decreased the salivary flow rate. The Na+, Cl- and K+ concentrations in the saliva were all significantly increased due to dysfunction of the ducts. Furthermore, Fam20c deficiency significantly increased BMP2 and BMP7 expression, decreased BMP4 expression, and attenuated the canonical and noncanonical BMP signaling pathways in the salivary glands. Collectively, the results of the present study demonstrate that FAM20C is a key regulator of acinar and duct structure and duct maturation and provide a novel avenue for investigating novel therapeutic targets for oral diseases including xerostomia.
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Células Acinares/patologia , Proteínas de Ligação ao Cálcio/deficiência , Proteínas da Matriz Extracelular/deficiência , Glândulas Salivares/patologia , Células Acinares/metabolismo , Células Acinares/ultraestrutura , Animais , Proteína Morfogenética Óssea 4/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Camundongos Knockout , Reprodutibilidade dos Testes , Saliva/metabolismo , Glândulas Salivares/metabolismo , Glândulas Salivares/ultraestrutura , Salivação , Transdução de Sinais , Glândula Submandibular/patologiaRESUMO
BACKGROUND: Oral tongue squamous cell carcinoma (OTSCC) is an aggressive cancer which has high mortality rates. HOXA transcript at the distal tip (HOTTIP) is a lncRNA that can be used as a prognostic marker in multiple carcinomas. The expression of HOTTIP is found to be elevated in OTSCC tissues, and such elevation is correlated with poor prognosis. However, its functional role in regulating the growth and metastasis of OTSCC cells remains elusive and requires further investigation. METHODS: HOTTIP-silenced OTSCC cells were established by inhibiting HOTTIP expression via its exclusive shRNA. Whether HOTTIP knockdown affected the aggressive tumor behaviors of OTSCC cells was investigated in vitro and in vivo. RESULTS: We found that HOTTIP shRNA restrained the cell proliferation and arrested the cell cycle at G1 phase in TSCCA and TCA8113 cells. The expression levels of cyclins B, D1, and E were downregulated in HOTTIP-silenced cells. HOTTIP silencing suppressed the growth of xenograft tumors. Moreover, the silencing of HOTTIP triggered apoptosis in TSCCA and TCA8113 cells and altered the expression of a group of apoptosis-related molecules: downregulated Bcl-2, upregulated Bax, and enhanced the cleavage of caspase 3 and PARP. Knockdown of HOTTIP also suppressed the migration, invasion, and epithelial-mesenchymal transition (EMT) of both TSCCA and TCA8113 cell lines. CONCLUSION: Our findings suggest that HOTTIP is required by the OTSCC cells to maintain their growth and metastasis in vitro. It may serve as a promising potential candidate for OTSCC therapy.
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Human dental pulp stem cells (hDPSCs) possess selfrenewal and osteogenic differentiation properties, and have been used for orofacial bone regeneration and periodontal treatment. Aspirin has been demonstrated to enhance the regeneration of bone marrow mesenchymal stem cells (MSCs); however, the impact of aspirin on the osteogenic differentiation of hDPSCs remains unknown. In the present study, hDPSCs were characterized by flow cytometry, while their clonogenic potential and multipotency were assessed using alizarin red, Oil red O and alcian blue staining. The effect of aspirin on hDPSC viability was assessed using Cell Counting Kit8 assay. Osteogenic capacity was examined by alkaline phosphatase activity, alizarin red staining, reverse transcriptionpolymerase chain reaction and western blotting. Furthermore, in vivo cranial defects were established in SpragueDawley rats to evaluate the effect of aspirin on hDPSCbased bone regeneration. Anorganic bovine bone was used as a bone replacement material and as the carrier for hDPSCs. New bone formation was observed through radiographic and histological analysis. The study demonstrated that hDPSCs expressed MSC markers and possessed multipotency in vitro. Aspirin was nontoxic to hDPSCs at a concentration of ≤100 µg/ml and enhanced the osteogenesis of hDPSCs in vitro. Aspirin significantly increased hDPSCbased bone formation in the rat cranial defect model at 8 or 12 weeks postimplantation (P<0.05). The data suggested that aspirin promotes the osteogenic potential of hDPSCs in vitro and in vivo. Overall, the present study indicated that aspirin improves the bone regeneration capacity of hDPSCs.
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Aspirina/farmacologia , Diferenciação Celular/efeitos dos fármacos , Polpa Dentária/citologia , Adolescente , Fosfatase Alcalina/metabolismo , Animais , Western Blotting , Feminino , Citometria de Fluxo , Humanos , Masculino , Microscopia Eletrônica , Osteogênese/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células-Tronco/citologia , Células-Tronco/efeitos dos fármacosRESUMO
Oral squamous cell carcinoma (OSCC), one of the 10 most common types of neoplasms in the US, constitutes ~90% of all cases of oral malignancies. Chemoresistance and metastasis are difficult to avoid during the course of treatment, leading to a poor prognosis and a high mortality rate for patients with OSCC. Autophagy, a critical conserved cellular process, has been reported to be highly associated with the regulation of chemoresistance and metastasis of cancer cells. The present study investigated the role of karyopherin α2 (KPNA2), a member of the importin α family, which may serve an important role in p53 nucleocytoplasmic transport in the process of OSCC autophagy. In the CAL27, SCC15 and Tca8113 OSCC cell lines, we observed that the downregulation of KPNA2 suppressed cell migration and cisplatin resistance, using woundhealing, Transwell and CCK8 assays. Additionally, the results of western blot analysis and transmission electron microscopy (TEM) analysis indicated that the knockdown of KPNA2 inhibited autophagy. We confirmed that the inhibition of autophagy with antiautophagy agents decreased the migration and cisplatin resistance of OSCC cells. We hypothesized that the suppression of cell migration and cisplatin resistance induced by KPNA2 knockdown may be associated with the inhibition of autophagy. To identify the underlying mechanism, further experiments determined that KPNA2 affects the level of autophagy via regulating the p53 nuclear import. Thus, the present study demonstrated that the function of KPNA2 in the process of autophagy may be p53dependent, and by regulating the translocation of p53, KPNA2 can support autophagy to promote the chemoresistance and metastasis of OSCC cells.
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Carcinoma de Células Escamosas/tratamento farmacológico , Neoplasias Bucais/tratamento farmacológico , Proteína Supressora de Tumor p53/genética , alfa Carioferinas/genética , Transporte Ativo do Núcleo Celular/genética , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Autofagia/genética , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Linhagem da Célula/genética , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Cisplatino/farmacologia , Resistencia a Medicamentos Antineoplásicos/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Humanos , Neoplasias Bucais/genética , Neoplasias Bucais/patologia , Transporte Proteico/genética , alfa Carioferinas/antagonistas & inibidoresRESUMO
Aspirin has positive effects on bone marrow mesenchymal stem cells (BMSCs) osteogenic differentiation. However, researchers did not give much thought to its effect on BMSCs adipogenic differentiation. Here, we analyzed the effect of aspirin on the BMSCs adipogenic differentiation. To detect whether the effect of aspirin on the adipogenic differentiation of BMSCs is associated with the disturbed epigenetic modification, the expression of histone deacetylases (HDACs), activity of HDACs and HAT, global histone H3 acetylation and H3k9 acetylation alterations were investigated. Moreover, to further explore and understand the binding mode between aspirin and HDACs, an attempt was made to identify the interaction between aspirin and the HDACs with the aid of in silico docking study. The results showed that aspirin could induce inhibition of BMSCs adipogenesis. The level of HDAC activity, global histone H3 acetylation, and H3k9 acetylation were all down regulated during adipogenic differentiation, and aspirin can reverse these decreases. Furthermore, the HDAC isoforms have different expression patterns in those progresses. The expression of HDAC9 was increased in a does-dependent manner when aspirin was introduced during BMSCs adipogenic differentiation. Docking study showed that high affinity of HDAC9 to aspirin was existed, suggesting that HDAC9 may has an important role in the process of aspirin-induced suppression of adipogenesis. Further studies are needed to define the intricate mechanisms of the HDAC isoforms, and all of these enable us to understand aspirin and its efficacy of inhibition of adipogenic differentiation and pave the way to aspirin clinical using for the tissue regenerating.
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Adipogenia , Tecido Adiposo/citologia , Aspirina/farmacologia , Células da Medula Óssea/citologia , Diferenciação Celular/efeitos dos fármacos , Epigênese Genética , Células-Tronco Mesenquimais/citologia , Acetilação , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/metabolismo , Sequência de Aminoácidos , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Apoptose , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Proliferação de Células , Células Cultivadas , Histona Desacetilases/química , Histonas/genética , Histonas/metabolismo , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Simulação de Acoplamento Molecular , Conformação Proteica , Ratos , Ratos Sprague-Dawley , Homologia de SequênciaRESUMO
After Peripheral nerve injuries (PNI), many complicated pathophysiologic processes will happen. A global view of functional changes following PNI is essential for the looking for the adequate therapeutic approaches. In this study, we performed an in-depth analysis on the temporal expression profiles after sciatic nerve injury by bioinformatic methods, including (1) cluster analysis of the samples; (2) identification of gene co-expression modules(CEMs) correlated with the time points; (3) analysis of differentially expressed genes at each time point (DEGs-ET); (4) analysis of differentially expressed genes varying over time (DEGs-OT); (5) creating Pairwise Correlation Plot for the samples; (6) Time Series Regression Analysis; (7) Determining the pathway, GO (gene ontology) and drug by enrichment analysis. We found that at a 3 h "window period" some specific gene expression may exist after PNI, and responses to lipopolysaccharide (LPS) and TNF signaling pathway may play important roles, suggesting that the inflammatory microenvironment exists after PNI. We also found that troglitazone was closely associated with the change of gene expression after PNI. Therefore, the further evaluation of the precise mechanism of troglitazone on PNI is needed and it may contribute to the development of new drugs for patients with PNI.
Assuntos
Traumatismos dos Nervos Periféricos/genética , Neuropatia Ciática/genética , Transcriptoma , Análise por Conglomerados , Biologia Computacional/métodos , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica , Ontologia Genética , Redes Reguladoras de Genes , Humanos , Anotação de Sequência Molecular , Traumatismos dos Nervos Periféricos/metabolismo , Neuropatia Ciática/metabolismo , Transdução de SinaisRESUMO
Exacerbations of oral lichen planus (OLP) have been linked to the periods of psychological stress, anxiety and depression. The specific mechanism of the interaction is unclear. The aim of this study was to explore the candidate genes or molecules that play important roles in the interaction between OLP and depression. The BITOLA system was used to search all intermediate concepts relevant to the "Gene or Gene Product" for OLP and depression, and the gene expression data and tissue-specific gene data along with manual checking were then employed to filter the intermediate concepts. Finally, two genes (NCAM1, neural cell adhesion molecule 1; CD4, CD4 molecule) passed the follow-up inspection. By using the text mining can formulate a new hypothesis: NCAM1 and CD4 were identified as involved or potentially involved in the interaction between OLP and depression. These results offer a new clue for the experimenters and hold promise for developing innovative therapeutic strategies for these two diseases.
Assuntos
Encéfalo/metabolismo , Antígenos CD4/metabolismo , Antígeno CD56/metabolismo , Depressão/metabolismo , Depressão/patologia , Líquen Plano Bucal/metabolismo , Antígenos CD4/genética , Antígeno CD56/genética , Eletroencefalografia , Feminino , Humanos , Masculino , RNA Mensageiro/metabolismo , Transcriptoma/fisiologiaRESUMO
Hypoxia regulates a number of cell biological processes, including cell survival, development and differentiation. Deferoxamine (DFO), an oral chelator for blood transfusion patients, has been demonstrated to induce hypoxia and is frequently used as a hypoxiamimicking agent. The purpose of the present study was to investigate the influence of DFO on the proliferation, migration and osteogenic differentiation of human periodontal ligament cells (hPDLCs). The effects of DFO on hPDLC viability and migration were measured using an MTT and wound healing assay. To characterize the hypoxia microenvironment, the expression of hypoxiainducible factor1α (HIF1α) in hPDLCs treated with DFO was quantified using the reverse transcriptionquantitative polymerase chain reaction (RTqPCR). Subsequently, the osteogenic differentiation potential of DFO was determined by RTqPCR of the mRNA of osteogenic markers (runtrelated transcription factor 2 [Runx2], osteopontin [OPN] and collagen type I [Col1]). The alkaline phosphatase activity and mineral deposition were analyzed using alizarin red S staining. The MTT and wound healing assays demonstrated that lowconcentrations of DFO had little impact on hPDLC viability and migration 48 h into the treatment. DFO upregulated the expression of hPDLC genes specific for osteogenic differentiation: HIF1α, Runx2, OPN and Col1. Furthermore, formation of mineralized nodules was enhanced by DFO. The present study suggests that DFO provided favorable culture conditions to promote the osteogenic differentiation and mineralization of hPDLCs. The mechanism underlying these alterations remains to be elucidated.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Desferroxamina/farmacologia , Osteogênese/efeitos dos fármacos , Fosfatase Alcalina/metabolismo , Biomarcadores , Diferenciação Celular/genética , Sobrevivência Celular/efeitos dos fármacos , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Osteogênese/genética , Ligamento Periodontal/citologia , CicatrizaçãoRESUMO
Cirrhosis is the terminal stage of hepatic diseases and is prone to develop into hepatocyte carcinoma. Increasing evidence suggests that the transplantation of dental pulp stem cells (DPSCs) may promote recovery from cirrhosis, but the key regulatory mechanisms involved remain to be determined. In this study, we overexpressed human hepatocyte growth factor (hHGF) in primary rat DPSCs and evaluated the effects of HGF overexpression on the biological behaviors and therapeutic efficacy of grafted DPSCs in cirrhosis. Liver cirrhosis was induced via the intraperitoneal injection of CCl4 twice weekly for 12 weeks and was verified through histopathological and serological assays. HGF was overexpressed in DPSCs via transduction with a hHGF-lentiviral vector and confirmed based on the elevated expression and secretion of HGF. The HGF-overexpressing DPSCs were transplanted into rats intravenously. The HGF-overexpressing DPSCs showed increased survival and hepatogenic differentiation in host liver tissue at 6 weeks after grafting. They also exhibited a significantly greater repair potential in relation to cirrhosis pathology and impaired liver function than did DPSCs expressing HGF at physiological levels. Our study may provide an experimental basis for the development of novel methods for the treatment of liver cirrhosis in clinical practice.