ABSTRACT
MicroRNA (miR) is a class of highly conserved non-coding single-stranded RNA widely existing in mammals, which can negatively regulate the expression of targeting genes after transcription. As a key regulator, miR negatively regulates the expression of the targeting genes and disrupts important molecular signaling pathways, leading to the imbalance of multiple pathways such as tissue repair and inflammation involved in the fibrotic process. Among them, miR-15a/16 can participate in regulating and controlling the fibrotic process of various organs, including liver, lung, heart, kidney and other fibrotic diseases by acting on cell proliferation and transformation, extracellular matrix proteins production and degradation, inflammation and other important cell functions. It has potential diagnostic and therapeutic value. Clarifying the biological function of miR-15a/16 and its mechanism for action and therapeutic application prospects in various fibrotic lesions are of great significance for the molecular targeted treatment of fibrotic diseases.
Subject(s)
Humans , Fibrosis/genetics , MicroRNAs/genetics , Signal Transduction , InflammationABSTRACT
Abstract Objective Osteoarthritis is a condition characterized by articular cartilage degradation. The increased expression of β1,4-Galactosyltransferase-I (β1,4-GalT-I) in the articular cartilage of osteoarthritis patients was related to an inflammatory response. The aim of this study was to elucidate the role of β1,4-GalT-I in osteoarthritis. This study aimed to determine the function of 1,4-GalT-I in osteoarthritis. Methods The osteoarthritis mouse model with the destabilization of the medial meniscus was established by microsurgical technique. Pathological changes in articular cartilage were observed by hematoxylin and eosin staining and safranin O-fast green staining. Quantitative real-time polymerase chain reaction, western blot, and enzyme-linked immunosorbent assays were used to observe mRNA and protein expression, respectively. RNA interactions were verified by a luciferase reporter assay. SA-β-Gal staining was used to assess chondrocyte senescence. Immunofluorescence staining was conducted to observe the localization of Nuclear Factor-kappaB (NF-κB). Results β1,4-GalT-I and microRNA-15a (miR-15a) show high and low expression in the articular cartilage of osteoarthritis, respectively. MiR-15a inhibits the mRNA translation of β1,4-GalT-I. β1,4-GalT-I promotes extracellular matrix degradation, senescence, and NF-κB activation in IL-1β-stimulated chondrocytes, which can be reversed by overexpression of miR-15a. Intra-articular injection of microRNA-15a ameliorates cartilage degeneration by inhibiting β1,4-GalT-I and phosphorylation of NF-κB in vivo. Conclusion The authors clarified that the miR-15a/β1,4-GalT-I axis inhibits the phosphorylation of NF-κB thereby inhibiting extracellular matrix degradation and senescence in chondrocytes to alleviate cartilage degeneration in osteoarthritis. MiR-15a and β1,4-GalT-I may serve as potentially effective targets for the future treatment of osteoarthritis.
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Objective:To investigate the correlation between serum microRNA (miR)-15a-5p and prognosis, neoadjuvant chemotherapy (NAC) response in patients with locally advanced gastric cancer (LAGC).Methods:The clinical data of 122 patients with LAGC who underwent surgery after NAC in Eastern Theater Air Force Hospital of the Chinese People′s Liberation Army from May 2016 to April 2020 were analyzed retrospectively. The general clinical data and laboratory examination results of the patients were recorded. The expression level of serum miR-15a-5p was detected by real-time fluorescence quantitative polymerase chain reaction, and the relationship between the expression of miR-15a-5p and different clinical characteristics in patients with LAGC was analyzed. The pathological response was evaluated by Becker tumor regression grading, in which patients with grade 1a, 1b and 2 were sensitive group and patients with grade 3 were resistant group.Results:The patients with LAGC were divided into high expression (>1.038) and low expression (≤1.038) according to the median miR-15a-5p of 1.038 with 61 cases each. The expression level of serum miR-15a-5p was related to the preference for spicy food, endoscopic ultrasonography (EUS)-T stage and EUS-N stage ( P<0.01 or <0.05). According to the evaluation result of pathological reaction, there were 47 cases in resistance group and 74 cases in sensitive group. The serum miR-15a-5p in resistance group was significantly higher than that in sensitive group: 1.69 (1.39, 1.97) vs. 0.99 (0.96, 1.02), and there was statistical difference ( Z =-8.55, P<0.01). The receiver operating characteristic curve analysis result showed that the area under the curve of serum miR-15a-5p predicting NAC response was 0.959 (95% CI 0.929 to 0.990), the optimal cut-off value was 1.049, the sensitivity was 100.0%, and the specificity was 85.1%. Multivariate Logistic regression analysis result showed that miR-15a-5p was an independent risk factor for NAC response in patients with LAGC ( HR = 1 880.840, 95% CI 123.510 to 28 641.846, P<0.01). Kaplan-Meier survival curve analysis result showed that the median overall survival time and median progression free survival time in patients with high expression of miR-15a-5p were significantly shorter than those in patients with low expression of miR-15a-5p (19 months vs. 62 months and 12 months vs. 51 months), and there were statistical differences (log-rank χ2 = 41.99 and 61.97, P<0.01); the 10-year overall survival rate and 10-year progression free survival rate in patients with high expression of miR-15a-5p were significantly lower than those in patients with low expression of miR-15a-5p (4.9% vs. 52.5% and 24.6% vs. 85.2%), and there were statistical differences (log-rank χ2 = 33.70 and 45.32, P<0.01). Multivariate Cox regression analysis result showed that R 0 resection and miR-15a-5p were the independent risk factors affecting the overall survival time and progression free survival time in patients with LAGC (overall survival time: HR = 1.945 and 3.487, 95% CI 1.033 to 3.660 and 2.112 to 5.759, P<0.05 or <0.01; progression free survival time: HR = 2.427 and 6.335, 95% CI 1.069 to 5.510 and 3.341 to 12.013, P<0.05 or <0.01). Conclusions:The increase of serum miR-15a-5p level is related to NAC response and poor prognosis in patients with LAGC. It can be used as a reliable biomarker to predict the prognosis and NAC response of LAGC.
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Objective:To investigate the effect of microRNA-15a (miR-15a) on the anti-oxidative stress ability of human lens epithelial cells (LECs) induced by high glucose and its possible mechanism.Methods:The anterior lens capsule specimens from patients with diabetic cataract (DC) and healthy donors were collected.The expressions of miR-15a and silent information regulator 1 (SIRT1) in the specimens were detected by real-time quantitative PCR (RT-qPCR) and Western blot, respectively.The human lens epithelial cell line HLEB-3 cells were cultured with 0, 10, 20, or 50 mmol/L glucose for 24 hours.The expression of miR-15a in the cells was detected by RT-qPCR.The expressions of SIRT1, forkhead transcription factor 3a (FOXO3a), and p53 proteins in the cells were determined by Western blot.The cell apoptosis was assayed by flow cytometry.The endogenous reactive oxygen species (ROS) content in the cells was measured by 2, 7-dichlorodihydrofluorescein diacetate (DCFH-DA). The total antioxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) activity, and malondialdehyde (MDA) activities in the cells were identified.HLEB-3 cells were transfected with miR-15a control or miR-15a inhibitor, then incubated with 50 mmol/L glucose for 24 hours.Cell apoptosis of the transfected cells was detected by flow cytometry.The endogenous ROS expression in the transfected cells was determined by DCFH-DA.The T-AOC, SOD, and GSH-Px activities as well as MDA concentration were measured.The relationship between miR-15a and SIRT1 was verified by dual-luciferase reporter assay.The SIRT1, FOXO3a, and p53 protein expressions in the transfected cells were detected by Western blot.This study was approved by an Ethics Committee of the Second Affiliated Hospital of Zhengzhou University (No.ZDEFY201803160023). Written informed consent was obtained from each subject.Results:The relative expression of miR-15a in normal lens anterior capsule tissue was 0.21±0.02, which was lower than 0.96±0.10 in lens anterior capsule tissue of DC patients, and the difference was statistically significant ( t=12.231, P<0.001). The relative expression of SIRT1 in the anterior lens capsule tissue was 0.89±0.09, which was higher than 0.31±0.05 in the anterior lens capsule tissue of DC patients, showing a statistically significant difference ( t=8.964, P<0.001). With the increase of glucose concentration, the relative expression of miR-15a, FOXO3a, and p53 in cells increased, and the relative expression of SIRT1 decreased; the apoptosis rate of cells increased; the ROS content and MDA concentration increased; the activities of T-AOC, SOD and GSH-Px decreased, and the differences were statistically significant (all at P<0.05). The apoptosis rate, ROS content, and MDA concentrations were higher in miR-15a control group than miR-15a inhibitor group, and the activities of T-AOC, SOD, and GSH-Px were lower in miR-15a control group than miR-15a inhibitor group, with statistically significant differences (all at P<0.05). The luciferase activity of the SIRT1-3'-untranslated region (UTR)-wild type (WT) reporter gene in miR-15a control group was significantly lower than that in miR-15a inhibitor group, and the difference was statistically significant ( t=5.978, P=0.004). No significant difference was found in the luciferase activity of the SIRT1-3'-UTR-mutant type (MUT) reporter gene ( t=0.432, P=0.688). The relative expressions of FOXO3a and p53 proteins were significantly higher in miR-15a control group than miR-15a inhibitor group, and the relative expression of SIRT1 protein was significantly lower in miR-15a control group than miR-15a inhibitor group, showing statistically significant differences (all at P<0.05). Conclusions:miR-15a can inhibit the anti-oxidative stress damage ability of LECs induced by high glucose, which may be achieved by inhibiting the expression of SIRT1 to up-regulate the activities of FOXO3a and p53, and aggravating apoptosis.
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The occurrence of thyroid gland is easily affected by iodine deficiency, the enzyme defect, drugs, autoimmune and other factors. The increased incidence of thyroid cancer year by year has threatened the health of the human, which requires to study thyroid gland cancer invasion and metastasis, investigate the molecular mechanism of cancer metastasis, search the differential molecular expression gene, predict the metastatic biomarkers and the intervening treatment target molecule, in order to improve the cure rate and the survival rate. This paper reviews the possible role of long chain non-coding RP11-23J9.4-miRNA-15a-axis inhibiting factor 2-Wnt pathway in the occurrence, development and dedifferentiation of thyroid cancer.