miRNA-382-5p Carried by Extracellular Vesicles in Osteoarthritis Reduces Cell Viability and Proliferation, and Promotes Cell Apoptosis by Targeting PTEN.

IF 2.6 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Hanyu Lu, Yixin Yang, Shuanji Ou, Yong Qi, Guitao Li, Hebei He, Fanglian Lu, Wenjun Li, Hongtao Sun
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引用次数: 0

Abstract

The objective of the study was to identify extracellular vesicle (EV) microRNAs (miRNAs) that play important roles in knee osteoarthritis (OA). Models of knee OA were surgically induced in nine male Sprague-Dawley rats. Tissue samples were collected at 0 weeks (Control), 6 weeks (6 weeks), and 12 weeks (12 weeks). The EVs were isolated and analyzed for size. Various biomarkers, including recombinant tetraspanin 30 cluster of differentiation (CD)63 and CD9 were detected. An Agilent array was used to screen for differentially expressed (DE) miRNAs. The levels of DE miRNAs and their target mRNAs were evaluated by quantitative reverse transcription-polymerase chain reaction and western blotting. The viability, proliferation, and apoptosis of lipopolysaccharide (LPS)-induced human synovial cells (HSCs) were examined by using Cell Counting Kit-8, EdU (5-ethynyl-2'-deoxyuridine), and TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling) assays, respectively. The OA model rats had significantly increased levels of inflammatory activity, damaged cells, and rough articular cartilage when compared with rats in the control group. The EVs from the model rats appeared as round vesicle-like structures with a mean diameter of ∼145 nm. Five miRNAs that showed gradual increases in the model rats were selected for further analysis; those miRNAs included miR-127-3p, miR-132-3p, miR-141-3p, miR-345-5p, and miR-382-5p. miR-382-5p was found to reduce the viability and proliferation and promote the apoptosis of LPS-induced HSCs. Moreover, phosphatase and tensin homolog deleted on chromosome 10 (PTEN) was negatively regulated by miR-382-5p. Our findings revealed that EVs produced by the OA rats contained miR-382-5p, which might reduce cell viability and proliferation, and promote cell apoptosis by targeting PTEN.

骨关节炎细胞外小泡携带的miRNA-382-5p通过靶向PTEN降低细胞活力和增殖,促进细胞凋亡
该研究的目的是鉴定在膝关节骨关节炎(OA)中起重要作用的细胞外囊泡(EV) microRNAs (miRNAs)。9只雄性Sprague-Dawley大鼠手术诱导膝关节OA模型。分别于0周(对照组)、6周(6周)和12周(12周)采集组织样本。分离ev并对其大小进行分析。检测多种生物标志物,包括重组tetraspanin 30 cluster of differentiation (CD)63和CD9。使用Agilent阵列筛选差异表达(DE) mirna。通过定量逆转录-聚合酶链反应和western blotting检测DE mirna及其靶mrna的水平。采用细胞计数试剂盒-8、EdU(5-乙基-2′-脱氧尿苷)和TUNEL(末端脱氧核苷酸转移酶介导的dUTP镍端标记法)检测脂多糖(LPS)诱导的人滑膜细胞(hsc)的活力、增殖和凋亡。与对照组大鼠相比,OA模型大鼠的炎症活性、细胞损伤和关节软骨粗糙程度显著增加。模型大鼠的EVs呈圆形囊泡状结构,平均直径约145 nm。选择5个在模型大鼠中逐渐增加的mirna进行进一步分析;这些mirna包括miR-127-3p、miR-132-3p、miR-141-3p、miR-345-5p和miR-382-5p。发现miR-382-5p可降低脂多糖诱导的hsc的活力和增殖,促进其凋亡。此外,10号染色体上缺失的磷酸酶和紧张素同源物(PTEN)受到miR-382-5p的负调控。我们的研究结果表明,OA大鼠产生的ev中含有miR-382-5p, miR-382-5p可能通过靶向PTEN降低细胞活力和增殖,促进细胞凋亡。
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来源期刊
DNA and cell biology
DNA and cell biology 生物-生化与分子生物学
CiteScore
6.60
自引率
0.00%
发文量
93
审稿时长
1.5 months
期刊介绍: DNA and Cell Biology delivers authoritative, peer-reviewed research on all aspects of molecular and cellular biology, with a unique focus on combining mechanistic and clinical studies to drive the field forward. DNA and Cell Biology coverage includes: Gene Structure, Function, and Regulation Gene regulation Molecular mechanisms of cell activation Mechanisms of transcriptional, translational, or epigenetic control of gene expression Molecular Medicine Molecular pathogenesis Genetic approaches to cancer and autoimmune diseases Translational studies in cell and molecular biology Cellular Organelles Autophagy Apoptosis P bodies Peroxisosomes Protein Biosynthesis and Degradation Regulation of protein synthesis Post-translational modifications Control of degradation Cell-Autonomous Inflammation and Host Cell Response to Infection Responses to cytokines and other physiological mediators Evasive pathways of pathogens.
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