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Revascularization of coronary chronic total occlusion (CTO) still remains controversial. The factors that impact collateral circulation and myocardial perfusion are of interest. Circular RNA (circRNA) has been shown to regulate the process of angiogenesis. However, the effects of circ-membrane-bound O-acyltransferase domain containing 2 (circ-MBOAT2) on angiogenesis in patients with CTO were unclear. In this study, we evaluated circulating circRNAs and miRNAs in patients with CTO and stable coronary artery disease using high-throughput sequencing. Another cohort of patients were selected to verify the expressions of circ-MBOAT2 and miR-495. The role and mechanism of circ-MBOAT2 in the process of angiogenesis were explored through in vitro and vivo studies. Finally, we came back to a clinical perspective and investigated whether circ-MBOAT2 and miR-495 were associated with the improvement of myocardial perfusion evaluated by single-photon emission computed tomography (SPECT). We found that the expression of circ-MBOAT2 was significantly up-regulated while miR-495 was significantly down-regulated in patients with CTO. The expression of circ-MBOAT2 was negatively correlated with miR-495 in patients with CTO. In an in vitro study, we found that circ-MBOAT2 promoted tube formation and cell migration via the miR-495/NOTCH1 axis in endothelial cells. In an in vivo study, we showed that the inhibition of miR-495 caused the increase in collateral formation in mice after hindlimb ischemia. In a human study, we showed the expressions of circ-MBOAT2 and miR-495 were associated with myocardial perfusion improvement after revascularization of CTO. In conclusion, circ-MBOAT2 regulates angiogenesis via the miR-495/NOTCH1 axis and associates with myocardial perfusion in patients with CTO. Our findings suggest that circ-MBOAT2 and miR-495 may be potential therapeutic targets and prognostic factors for patients with CTO. [ABSTRACT FROM AUTHOR]

miR-495 is aberrantly expressed and affects the progression of inflammation in various diseases. However, the mechanisms of miR-495 in bovine endometritis remain largely unknown. This study investigated the mechanism of miR-495 in lipopolysaccharide (LPS)-induced bovine endometritis and pyroptosis and found that miR-495 inhibits NLRP3 inflammasome activation and inflammatory immune responses in endometritis tissue and cell models. Bovine endometrial epithelial cells (BENDs) were treated with 10 μg/mL LPS to establish a cell inflammatory model. LPS stimulation activated the NLRP3 inflammasome and elevated the expression of proinflammatory factors in BEND cells. In addition, pyroptosis and methylation-dependent inhibition of miR-495 was discovered in LPS-exposed BENDs. Furthermore, overexpression of miR-495 inhibited activation of the NLRP3 inflammasome in vitro and vivo. Collectively, our data demonstrate that miR-495 can attenuate activation of the NLRP3 inflammasome to protect against pyroptosis and bovine endometritis, which provides novel therapeutic targets for bovine endometritis and other inflammatory diseases. [Display omitted] • Overexpression of miR-495 could inhibit activation of the NLRP3 inflammasomein bovine endometritis. • Methylation of miR-495 promoter suppresses the expression of miR-495. • Revealed the anti-inflammatory and anti-pyroptosis effects of miR-495–3p in vivo. [ABSTRACT FROM AUTHOR]

Background: Diabetic Kidney Disease (DKD) is a significant challenge in healthcare. However, there are currently no reliable biomarkers for renal impairment diagnosis, prognosis, or staging in DKD patients. CircRNAs and microRNAs have emerged as noninvasive and efficient biomarkers. Methods: We explored Cannabinoid receptor 1 (CNR1), C reactive protein (CRP), hsa_circ_ 0000146 and 0000072, and hsa-miR-21 and 495 as diagnostic biomarkers in DKD. The serum concentrations of CRP and CNR1 were measured using ELISA. Rt-qPCR was used to evaluate the expression levels of CNR1, circRNAs, and miRNAs in 55 controls, 55 type 2 diabetes mellitus patients, and 55 DKD patients. Their diagnostic value was determined by their ROC curve. KEGG pathway was used to predict the functional mechanism of the circRNA's target genes. Results: DKD patients exhibited a significant increase in CRP and CNR1 levels and the expression of miR-21 and 495. The expression levels of circ_0000146 and 0000072 decreased in DKD patients. ROC analysis revealed that circRNAs and miRNAs alone or CNR1 and CRP have significant diagnostic potential. The functional prediction results showed the involvement of hsa_circ_0000146 and 0000072 in various pathways that regulate DKD. Conclusions: Therefore, the examined circRNAs and miRNAs may represent a novel noninvasive biomarker for diagnosing and staging DKD.

Background: Diabetic Kidney Disease (DKD) is a significant challenge in healthcare. However, there are currently no reliable biomarkers for renal impairment diagnosis, prognosis, or staging in DKD patients. CircRNAs and microRNAs have emerged as noninvasive and efficient biomarkers. Methods: We explored Cannabinoid receptor 1 (CNR1), C reactive protein (CRP), hsa_circ_ 0000146 and 0000072, and hsa-miR-21 and 495 as diagnostic biomarkers in DKD. The serum concentrations of CRP and CNR1 were measured using ELISA. Rt-qPCR was used to evaluate the expression levels of CNR1, circRNAs, and miRNAs in 55 controls, 55 type 2 diabetes mellitus patients, and 55 DKD patients. Their diagnostic value was determined by their ROC curve. KEGG pathway was used to predict the functional mechanism of the circRNA's target genes. Results: DKD patients exhibited a significant increase in CRP and CNR1 levels and the expression of miR-21 and 495. The expression levels of circ_0000146 and 0000072 decreased in DKD patients. ROC analysis revealed that circRNAs and miRNAs alone or CNR1 and CRP have significant diagnostic potential. The functional prediction results showed the involvement of hsa_circ_0000146 and 0000072 in various pathways that regulate DKD. Conclusions: Therefore, the examined circRNAs and miRNAs may represent a novel noninvasive biomarker for diagnosing and staging DKD. [ABSTRACT FROM AUTHOR]

Single-nucleotide polymorphisms (SNPs) of microRNAs (miRNAs) may alter miRNA transcription, maturation and target specificity, thus affecting stroke susceptibility. We aimed to investigate whether miR-200b and miR-495 SNPs may be associated with ischemic stroke (IS) risk and further explore underlying mechanisms including related genes and pathways. MiR-200b rs7549819 and miR-495 rs2281611 polymorphisms were genotyped among 712 large-artery atherosclerosis (LAA) stroke patients and 1,076 controls in a case–control study. Bioinformatic analyses were performed to explore potential association of miR-200b/495 with IS and to examine the effects of these two SNPs on miR-200b/495. Furthermore, we evaluated the association between these two SNPs and stroke using the public GWAS datasets. In our case–control study, rs7549819 was significantly associated with a decreased risk of LAA stroke (OR = 0.73, 95% CI = 0.58–0.92; p = 0.007), while rs2281611 had no significant association with LAA stroke risk. These results were consistent with the findings in East Asians from the GIGASTROKE study. Combined effects analysis revealed that individuals with 2–4 protective alleles (miR-200bC and miR-495 T) exhibited lower risk of LAA stroke than those with 0–1 variants (OR = 0.76, 95% CI = 0.61–0.96; p = 0.021). Bioinformatic analyses showed that miR-200b and miR-495 were significantly associated with genes and pathways related to IS pathogenesis, and rs7549819 and rs2281611 markedly influenced miRNA expression and structure. MiR-200b rs7549819 polymorphism and the combined genotypes of miR-200b rs7549819 and miR-495 rs2281611 polymorphisms were associated with decreased risk of LAA stroke in Chinese population. [ABSTRACT FROM AUTHOR]

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Abstract Background Knowledge of stenosis in coronary arteries requires an understanding of the cellular and molecular processes that occur throughout the leukocyte rolling process. In this study, the roles of miR-125a-5p and miR-495-3p were investigated on the adhesion of endothelial cells (ECs) isolated from the human aorta. Methods Human primary endothelial cells were obtained from the aorta of people who had died of brain death. Whole blood was used to isolate the monocytes. The miR-125 and miR-495 were predicted and transfected into ECs using Poly Ethylene Imine (PEI). The expression levels of adhesion molecules and monocyte recruitment were identified by the RT-qPCR technique and Leukocyte-Endothelial Adhesion Assay kit, respectively. Results The ICAM-1, ICAM-2 and VCAM-1 expression levels decreased significantly in the miR-495/PEI-transfected ECs (P

Abstract Background Hypoxia is a hallmark of cancer and is associated with poor prognosis. However, the molecular mechanism by which hypoxia promotes tumor progression remains unclear. MicroRNAs dysregulation has been shown to play a critical role in the tumor and tumor microenvironment. Here, we investigated the roles of miR‐495 and miR‐5688 in human non‐small cell lung cancer (NSCLC) and their underlying mechanism. Methods The expression levels of miR‐495 and miR‐5688 in human NSCLC tissue specimens were measured by quantitative real‐time polymerase chain reaction (qRT‐PCR). Deferoxamine (DFO) was used to determine whether the regulation of miR‐495 and miR‐5688 under hypoxia was dependent on hypoxia‐inducible factor 1‐alpha (HIF‐1α). Furthermore, the functions of miR‐495 and miR‐5688 in tumor progression were evaluated using colony formation, 3‐(4,5‐dimethylthiazol‐2‐yl)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium (MTS), wound healing, transwell assays, and xenograft model. Two algorithms, PicTAR and Targetscan, were used to predict the target gene of these two miRNAs, and dual‐luciferase reporter assay was conducted to confirm the target. The unpaired two‐tailed t test, Pearson correlation analysis, and Fisher's exact probability test were performed for statistical analyses. Results Two miRNAs, miR‐495 and miR‐5688, were found to participate in NSCLC progression under hypoxia. They were down‐regulated in NSCLC tissues compared with normal tissues. We determined that hypoxia led to the down‐regulation of miR‐495 and miR‐5688 in NSCLC cells, which was independent of HIF‐1α and cellular metabolic energy. In addition, miR‐495 and miR‐5688 suppressed cell proliferation, migration, and invasion in vitro. The NSCLC xenograft model showed that miR‐495 and miR‐5688 inhibited tumor formation in vivo. Interestingly, we found that miR‐495 and miR‐5688 had the same target, interleukin‐11 (IL‐11). Recombinant human IL‐11 counteracted the effects of miR‐495 and miR‐5688 on NSCLC cells, suggesting that miR‐495 and miR‐5688 executed their tumor suppressive role by repressing IL‐11 expression. Conclusion We found that hypoxia down‐regulated the expression levels of miR‐495 and miR‐5688 in NSCLC to enhance IL‐11 expression and tumor progression, indicating that the miR‐495/miR‐5688/IL‐11 axis may serve as a therapeutic target and potential biomarker for NSCLC.

Abstract Pneumonia is an inflammatory disease with leading mortality rate in children. It has been well established that microRNAs (miRNAs) have been regarded as critical regulator in acute lung injury. We intended to explore the effect and underlying mechanism of miR‐495 on lipopolysaccharides (LPS)‐induced WI‐38 cells. Here, we first found that miR‐495 was downregulated in serum of patients with acute stage pneumonia. To establish cell model of acute pneumonia, WI‐38 cells were treated with 20 μg/mL LPS, and qRT‐PCR analysis also confirmed the downregulation of miR‐495 in LPS‐induced WI‐38 cells. Data from MTT (3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5‐diphenyltetrazolium bromide) and flow cytometry assays showed that the decreased cell viability and induced cell apoptosis by LPS treatment were also reversed by miR‐495 over‐expression. Moreover, miR‐495 inhibited expression of associated inflammatory factors, which were induced by LPS treatment. Second, ROCK1 (rho‐associated, coiled‐coil‐containing protein kinase 1) was identified as functional target gene of miR‐495, whose expression was decreased by miR‐495. Mechanically, combination of miR‐495 and ROCK1 over‐expression reversed the influence of miR‐495 on LPS‐induced inflammation, viability, and apoptosis. In conclusion, our findings indicated that miR‐495 inhibited LPS‐induced inflammation injury and apoptosis in WI‐38 cells via targeting ROCK1, which would shed light on therapeutic schedule in acute pneumonia.

Abstract Background MicroRNAs (miRNAs) play crucial roles in regulating eukaryotic gene expression. Recent studies indicated that aberrantly expressed miRNAs are involved in the pathogenesis of ankylosing spondylitis (AS). Indeed, hsa-miR-495-3p (miR-495) has been reported as an anti-oncogene in different cancers. However, the role of miR-495 in AS is still unknown. Methods In this study, quantitative real-time polymerase chain reaction (PCR) was used to detect the expression of miR-495 in the peripheral blood mononuclear cells (PBMCs), whole blood, and serum of patients with AS. Bisulfite-specific PCR sequencing and methylated DNA immunoprecipitation were used to detect the methylation in the promoter region of miR-495. To determine the influence of miR-495 expression on the target gene, programmed cell death 10 (PDCD10), dual luciferase reporter assays together with an adenoviral vector containing the miR-495 locus were used. Receiver operating characteristic (ROC) curves were used to evaluate the efficacy of miR-495 as a diagnostic biomarker of AS. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes pathway analysis, and western blotting were used to explore the potential role of miR-495 in AS pathogenesis and the mechanism by which it facilitates AS pathogenesis. Results miR-495 is down-regulated and the promoter region of miR-495 is highly methylated in AS. The expression of miR-495 is negatively associated with PDCD10 expression in both patients with AS and healthy controls. Further experiments showed that PDCD10 can be targeted by miR-495. The ROC curves of miR-495 suggested that it is a very specific and sensitive biomarker for AS diagnosis. Bioinformatics analysis and signal pathway studies indicated that miR-495 can down-regulate β-catenin and transforming growth factor-β1. Conclusions Our studies indicated that down-regulation of miR-495 can be used as a potential molecular marker for the diagnosis and treatment of AS, thus providing new insights into the role of miRNAs in AS pathology.

Background: Cancer resistance to chemotherapy is closely associated with changes in transporter systems. In this study, we investigated the possible regulation of 1 copper ion transporter (ATP7A; ATPase copper transporting alpha) by microRNA miR-495 and its implications in cisplatin resistance and angiogenesis in esophageal cancer. Methods: MiR-495 and ATP7A mRNA expression in clinical tissue samples and 2 cancer cell lines (Eca-109 and TE1) were detected by quantitative real-time polymerase chain reaction. The levels of miR-495 and ATP7A expression in Eca-109 and TE1 cells were increased by transfection with miR-495 mimics and ATP7A-overexpression vectors. Cell proliferation, apoptosis, and angiogenesis were assessed by CCK-8, flow cytometry, and tube formation assays, respectively. The levels of TNF-α and VEGF in cell culture supernatants were detected by enzyme linked immunosorbent assay, and in situ expression of NLRP3 was measured by immunofluorescence. The binding of miR-495 to ATP7A sequences was verified by dual luciferase reporter assays. Results: ATP7A expression was significantly increased, while miR-495 expression was decreased in the cancer tissues of esophageal cancer patients. MiR-495 mimics decreased the proliferation and promoted the apoptosis of cisplatin-resistant Eca-109 and TE1 cells. Furthermore, tube formation by human umbilical vein endothelial cells, TNF-α and VEGF secretion, and the levels of MRP1, ABCG1, ABCA1, and NLRP3 expression in cisplatin-resistant Eca-109 and TE1 cells were all reduced by miR-495 mimics. MiR-495 was shown to directly bind to ATP7A gene sequences to repress ATP7A expression in Eca-109 and TE1 cells. ATP7A overexpression substantially abrogated the changes in proliferation, apoptosis, angiogenesis, and above-mentioned gene expression in cisplatin-resistant Eca-109 and TE1 cells. Conclusions: MiR-495 suppressed cisplatin resistance and angiogenesis in esophageal cancer cells by targeting ATP7A gene expression. [ABSTRACT FROM AUTHOR]

Background: Knowledge of stenosis in coronary arteries requires an understanding of the cellular and molecular processes that occur throughout the leukocyte rolling process. In this study, the roles of miR-125a-5p and miR-495-3p were investigated on the adhesion of endothelial cells (ECs) isolated from the human aorta.Methods: Human primary endothelial cells were obtained from the aorta of people who had died of brain death. Whole blood was used to isolate the monocytes. The miR-125 and miR-495 were predicted and transfected into ECs using Poly Ethylene Imine (PEI). The expression levels of adhesion molecules and monocyte recruitment were identified by the RT-qPCR technique and Leukocyte-Endothelial Adhesion Assay kit, respectively.Results: The ICAM-1, ICAM-2 and VCAM-1 expression levels decreased significantly in the miR-495/PEI-transfected ECs (P < 0.05) while in the miR-125/PEI-transfected ECs only the ICAM-2 and ITGB-2 expression levels decreased significantly (P < 0.05) as compared to the miR-synthetic/PEI-transfected ECs. Furthermore, the monocyte adhesion was decreased in the miR-125 and miR-mix/PEI-transfected ECs as compared to the miR-synthetic/PEI-transfected ECs (P = 0.01 and P = 0.04, respectively).Conclusion: According to the findings, the efficient relations between miR-125 and adhesion molecules may be responsible for the inhibition of monocyte rolling. [ABSTRACT FROM AUTHOR]

Choriocarcinoma is a rare gestational trophoblastic tumour. Propofol is one of the commonly used intravenous anaesthetic agents and exerts an anti-tumour role in multiple cancers. However, the effect of propofol on choriocarcinoma has not been well explored. We investigated the effect of propofol on JEG-3 cells. The effects of propofol on cell proliferation and metastasis were determined. The expression of miR-495 was suppressed while Twist1 overexpressed. Cell viability, migration and invasion were respectively measured using CCK-8 assay and transwell migration/invasion assay. The expression of miR-495, AKT1 and Twist1 was detected using qRT-PCR. Western blot analysis was performed to assess the expression levels of EMT- and proliferation-related factors, and TGF-β pathway proteins. Propofol treatment significantly inhibited cell proliferation, migration and invasion, and reduced the releasing of EMT. miR-495 was up-regulated by profocol, and miR-495 inhibition reversed the effect of propofol on cell proliferation and metastasis in JEG-3 cells. Furthermore, propofol decreased the expression of AKT1 and Twist1 through up-regulating miR-495, and Twist1 participated in the regulation of propofol on TGF-β signalling inactivation in JEG-3 cells. This study demonstrates that propofol suppresses cell proliferation and metastasis through up-regulation of miR-495 and down-regulation of Twist1 in JEG3 cells.HighlightsPropofol inhibits proliferation and metastasis in JEG-3 cells;miR-495 is up-regulated by propofol in JEG-3 cells;miR-495 suppression blocks the effect of propofol in JEG-3 cells;Propofol decreases Twist1 expression through up-regulating miR-495;Propofol inhibits TGF-β signalling via regulation of Twist1.

Abstract Background Sepsis leads to severe inflammatory and cardiac dysfunction. This study aimed to explore the clinical value of miR-495 in sepsis, as well as its role in sepsis-induced inflammation and cardiac dysfunction. Methods 105 sepsis patients were recruited; receiver operating characteristic (ROC) curve was plotted to assess the diagnostic value of miR-495 in sepsis. A model of sepsis in rats was created via performing cecal ligation and puncture (CLP). After modeling, the cardiac function, including left ventricular systolic pressure (LVSP), left ventricular end diastolic pressure (LVEDP) and maximum rate of rise/fall of left ventricle pressure (± dp/dtmax), and serum cardiac troponin I (CTn-I), creative kinase isoenzyme MB (CK-MB) were detected. The blood cytokines levels including TNF-α, IL-6, IL-1β were also measured. Quantitative real-time PCR (qRT-PCR) was used for the measurement of the expression level of miR-495. Results MiR-495 was significantly downregulated in sepsis patients, especially patients who suffered from septic shock (SS). MiR-495 expression was negatively associated with Scr, WBC, CRP, PCT, APACHE II score and SOFA score. MiR-495 could distinguish patients with SS from non-SS patients. MiR-495 and SOFA score were better indictors for the occurrence of cardiac dysfunction in sepsis patients. In CLP-induced sepsis model. CLP rats experienced deterioration of LVSP, LVEDP, ± dp/dtmax, and had a rise in serum CTn-I, CK-MB, TNF-α, IL-6 and IL-1β, which were improved by miR-495 agomir injection. Conclusions MiR-495 might be a potential diagnostic biomarker for sepsis patients, and overexpression of miR-495 alleviated sepsis-induced inflammation and cardiac dysfunction.

This study aims to investigate the role of miR-495 in neuronal cell apoptosis after acute spinal cord injury (ASCI). The ASCI rat model was established and the Basso, Beattie, and Bresnahan (BBB) score was assessed. miR-495, PR domain containing 5 (PRDM5), and Bcl-2 expressions were measured by qRT-PCR or western blotting. Neuronal cell line PC-12 was subjected to hypoxia condition to simulate the in vitro ASCI model. PC-12 cell apoptosis was measured by flow cytometry, and the interaction between miR-495 and PRDM5 was confirmed by dual luciferase reporter assay. Results showed that BBB score was significantly decreased in ASCI rats compared with sham rats. miR-495 expression was down-regulated in spinal cord tissue of ASCI rats and hypoxia-induced PC-12 cells, and PRDM5 protein level was up-regulated in spinal cord tissue of ASCI rats and hypoxia-induced PC-12 cells. miR-495 overexpression could reduce apoptosis of PC-12 cells, and up-regulated anti-apoptosis protein Bcl-2 protein level. Moreover, PRDM5 was a target of miR-495, and mRNA and protein levels of PRDM5 were negatively regulated by miR-495. miR-495 overexpression could reduce the hypoxia-induced PC-12 cell apoptosis, while PRDM5 overexpression abolished this inhibiting effect. The agomir-495 was injected into ASCI rats, and Bcl-2 protein level and BBB score were increased, but the PRDM5 overexpression reversed these results. Overall, we concluded that miR-495 could inhibit neuronal cell apoptosis and relieve acute spinal cord injury through inhibiting PRDM5. [ABSTRACT FROM AUTHOR]

Excessive chondrocyte apoptosis is mostly responsible for the progression of osteoarthritis (OA). It has been shown that circular RNAs (circRNAs) are differentially expressed in OA cartilage and participate in various pathological processes during OA. Here, this study was designed to explore the effect and molecular mechanism of hsa_circ_0005567 on IL-1β-induced chondrocyte apoptosis. The results showed that hsa_circ_0005567 knockdown aggravated the IL-1β-induced chondrocyte apoptosis. In contrast, hsa_circ_0005567 overexpression attenuated the IL-1β-induced chondrocyte apoptosis, but this effect could be abrogated by 3-methyladenine (an inhibitor of autophagy), suggesting that hsa_circ_0005567 overexpression inhibited chondrocyte apoptosis by inducing autophagy. Furthermore, hsa_circ_0005567 competitively bound to miR-495 and derepressed the expression of ATG14, an early autophagy marker that was a direct target of miR-495. Moreover, both miR-495 mimic and ATG14 knockdown counteracted the autophagy-promoting and anti-apoptotic effects of hsa_circ_0005567 overexpression in IL-1β-treated chondrocytes. Taken together, hsa_circ_0005567 activates autophagy by regulating the miR-495/ATG14 axis and thereby suppresses IL-1β-induced chondrocyte apoptosis. These findings suggest that hsa_circ_0005567 may serve as a therapeutic target for the treatment of OA.

The aim of the present study was to investigate the expression of microRNA (miR)-495 and miR-326 in the peripheral blood of patients with rheumatoid arthritis (RA). A total of 107 RA patients, admitted to the Yidu Central Hospital of Weifang (Weifang, China) from February 2016 to February 2019, and 112 healthy subjects, who underwent physical examination during the same period, were selected as the research subjects for prospective analysis. The RA patients served as the study group and the healthy subjects as the control group. The expression levels of miR-495 and miR-326 in the peripheral blood of the two groups of subjects were compared. The association between miR-495 and miR-326 with RA clinical pathology and the diagnostic value of miR-495 and miR-326 for RA were analyzed. In the study group, miR-495 expression was significantly higher than that in the control group, and miR-326 expression was significantly lower than that in the control group (P<0.001). miR-495 and miR-326 combined diagnosis showed good predictive value for the occurrence of RA (P<0.001) and was closely related to RA clinical pathology (P<0.001). After treatment, miR-495 expression was significantly decreased in the study group, whereas miR-326 expression was significantly increased (P<0.001). Pearson's correlation coefficient analysis showed that rheumatoid factor (RF) was positively correlated with miR-495 expression and negatively correlated with miR-326 expression (P<0.001). In conclusion, miR-495 was highly expressed in patients with RA, whereas miR-326 was poorly expressed in RA patients, and the combined detection of miR-495 and miR-326 has good diagnostic value for RA. [ABSTRACT FROM AUTHOR]

The present study aimed to explore the clinical significance of miR-372 and miR-495 in acute myeloid leukemia (AML). Eighty-one AML patients (research group) admitted to the First Hospital of Lanzhou University from March 2012 to January 2014 were selected, and 60 healthy persons (control group) were selected. The expression levels of miR-372 and miR-495 in the peripheral blood of the subjects were detected by reverse transcriptase quantitative PCR, and their diagnostic and prognostic values in AML were analyzed. The miR-372 expression level in the peripheral blood of patients in the research group was significantly higher than that in the control group (P<0.05), and the miR-495 level was significantly lower than that in the control group (P<0.05). The area under the curve (AUC), sensitivity, and specificity of miR-372 combined with miR-495 in the diagnosis of AML were 0.925, 86.43, and 93.33% respectively. The 5-year survival rate of patients with high expression of miR-372 was lower than that of those with low expression of miR-372 (P<0.05), and the 5-year survival rate of patients with high expression of miR-495 was higher than that of those with low expression of miR-495 (P<0.05). miR-372 and miR-495 were independent risk factors for the prognosis and survival of AML patients. miR-372 expression increased in AML, while miR-495 decreased. miR-372 and miR-495 are effective indicators for the early diagnosis and prognosis of AML. [ABSTRACT FROM AUTHOR]

Pneumonia is an inflammatory disease with leading mortality rate in children. It has been well established that microRNAs (miRNAs) have been regarded as critical regulator in acute lung injury. We intended to explore the effect and underlying mechanism of miR‐495 on lipopolysaccharides (LPS)‐induced WI‐38 cells. Here, we first found that miR‐495 was downregulated in serum of patients with acute stage pneumonia. To establish cell model of acute pneumonia, WI‐38 cells were treated with 20 μg/mL LPS, and qRT‐PCR analysis also confirmed the downregulation of miR‐495 in LPS‐induced WI‐38 cells. Data from MTT (3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5‐diphenyltetrazolium bromide) and flow cytometry assays showed that the decreased cell viability and induced cell apoptosis by LPS treatment were also reversed by miR‐495 over‐expression. Moreover, miR‐495 inhibited expression of associated inflammatory factors, which were induced by LPS treatment. Second, ROCK1 (rho‐associated, coiled‐coil‐containing protein kinase 1) was identified as functional target gene of miR‐495, whose expression was decreased by miR‐495. Mechanically, combination of miR‐495 and ROCK1 over‐expression reversed the influence of miR‐495 on LPS‐induced inflammation, viability, and apoptosis. In conclusion, our findings indicated that miR‐495 inhibited LPS‐induced inflammation injury and apoptosis in WI‐38 cells via targeting ROCK1, which would shed light on therapeutic schedule in acute pneumonia. [ABSTRACT FROM AUTHOR]

Background: MicroRNAs (miRNAs) play crucial roles in regulating eukaryotic gene expression. Recent studies indicated that aberrantly expressed miRNAs are involved in the pathogenesis of ankylosing spondylitis (AS). Indeed, hsa-miR-495-3p (miR-495) has been reported as an anti-oncogene in different cancers. However, the role of miR-495 in AS is still unknown. Methods: In this study, quantitative real-time polymerase chain reaction (PCR) was used to detect the expression of miR-495 in the peripheral blood mononuclear cells (PBMCs), whole blood, and serum of patients with AS. Bisulfite-specific PCR sequencing and methylated DNA immunoprecipitation were used to detect the methylation in the promoter region of miR-495. To determine the influence of miR-495 expression on the target gene, programmed cell death 10 (PDCD10), dual luciferase reporter assays together with an adenoviral vector containing the miR-495 locus were used. Receiver operating characteristic (ROC) curves were used to evaluate the efficacy of miR-495 as a diagnostic biomarker of AS. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes pathway analysis, and western blotting were used to explore the potential role of miR-495 in AS pathogenesis and the mechanism by which it facilitates AS pathogenesis. Results: miR-495 is down-regulated and the promoter region of miR-495 is highly methylated in AS. The expression of miR-495 is negatively associated with PDCD10 expression in both patients with AS and healthy controls. Further experiments showed that PDCD10 can be targeted by miR-495. The ROC curves of miR-495 suggested that it is a very specific and sensitive biomarker for AS diagnosis. Bioinformatics analysis and signal pathway studies indicated that miR-495 can down-regulate β-catenin and transforming growth factor-β1. Conclusions: Our studies indicated that down-regulation of miR-495 can be used as a potential molecular marker for the diagnosis and treatment of AS, thus providing new insights into the role of miRNAs in AS pathology. [ABSTRACT FROM AUTHOR]

Non‐small‐cell lung cancer (NSCLC) remains the leading cause of cancer death worldwide. As a platinum‐based chemotherapeutic drug, cisplatin has been used for over 30 years in NSCLC treatment while its effects are diminished by drug resistance. Therefore, we aimed to study the potential role of UCA1 in the development of chemoresistance against cisplatin. Real‐time polymerase chain reaction, western‐blot analysis, and immunofluorescence were used to study the involvement of UCA1, miR‐495, and NRF2 in chemoresistance against cisplatin. 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay was performed to determine the effect of cisplatin on cell proliferation. Computational analysis and luciferase assay were carried out to explore the interaction among UCA1, miR‐495, and NRF2. The cisplatin‐R group exhibited lower levels of UCA1 and NRF2 expression but a higher level of miR‐495 expression than the cisplatin‐S group. The growth rate and half‐maximal inhibitory concentration of cellular dipeptidyl peptidase (cisplatinum) of the cisplatin‐R group were much higher than those in the cisplatin‐S group. MiR‐495 contained a complementary binding site of UCA1, and the luciferase activity of wild‐type UCA1 was significantly reduced after the transfection of miR‐495 mimics. MiR‐495 directly targeted the 3′‐untranslated region (3′‐UTR) of NRF2, and the luciferase activity of wild‐type NRF2 3′‐UTR was evidently inhibited by miR‐495 mimics. Finally, UCA1 and NRF2 expressions in the effective group were much lower than that in the ineffective group, along with a much higher level of miR‐495 expression. We suggested for the first time that high expression of UCA1 contributed to the development of chemoresistance to cisplatin through the UCA1/miR‐495/NRF2 signaling pathway. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) play crucial roles in skin pigmentation in animals. Rainbow trout (Oncorhynchus mykiss) is a key economic fish species worldwide, and skin color directly affects its economic value. However, the functions of miRNAs in rainbow trout skin pigmentation remain largely unknown. Herein, we overexpressed and silenced miR-495 in vitro and in vivo to investigate its functions. The analysis of spatial and temporal expression patterns suggested that miR-495 is a potential regulator during the process of skin pigmentation. In vitro, mc1r was validated as a direct target for miR-495 by dual-luciferase reporter assay, and overexpression of miR-495 significantly inhibited mc1r expression; in contrast, mc1r and its downstream gene mitf levels were markedly upregulated by decreased miR-495. In vivo, overexpressed miR-495 by injecting agomiR-495 led to a substantial decrease in the expression of mc1r and mitf in dorsal skin and liver, while the opposite results were obtained after miR-495 silencing by antagomiR-495. These findings suggested that miR-495 can target mc1r to regulate rainbow trout skin pigmentation, which provide a potential basis for using miRNAs as target drugs to treat pigmentation disorders and melanoma. [ABSTRACT FROM AUTHOR]

Background/Aims: More and more reports have shown that the dysregulation of miRNAs can contribute to the progression and metastasis of human cancers. Many studies have shown that the down-regulation of the miR-495 level occurs in a variety of cancers, including colorectal cancer (CRC). However, the precise molecular mechanisms of miR-495 in CRC have not been well clarified. In the current study, we investigated the biological functions and molecular mechanisms of miR-495 in CRC cell lines. Methods: qRT-PCR was used to determine the level of miR-495 in CRC cell lines and tissues. A miR-495 mimic and inhibitor were transfected into CRC cells, and the effects of miR-495 on the invasion and EMT were explored by qRT-PCR as well as transwell and Western blot assays. Meanwhile, luciferase assays were performed to validate Annexin A3 as a miR-495 target in CRC cells. Results: In our study, we found that miR-495 is down-regulated in CRC tissues and cell lines. Moreover, the low level of miR-495 was associated with increased expression of Annexin A3 in CRC tissues and cell lines. The invasion and EMT of CRC cells were suppressed by the overexpression of miR-495. However, the down-regulation of miR-495 promoted the invasion and EMT of CRC cells. Bioinformatics analysis predicted that Annexin A3 was a potential target gene of miR-495. Next, the luciferase reporter assay confirmed that miR-495 could directly target Annexin A3. Consistent with the effect of miR-495, the down-regulation of Annexin A3 by siRNA inhibited the invasion and EMT of CRC cells through the up-regulation of p53. The introduction of Annexin A3 in CRC cells partially blocked the effects of the miR-495 mimic. Conclusion: The introduction of miR-495 directly targeted Annexin A3 to inhibit the invasion and EMT of CRC cells by up-regulating p53, and the down-regulation of Annexin A3 was essential for inhibiting the invasion and EMT of CRC cells by overexpressing miR-495. Overall, the re-activation of the miR-495/Annexin A3/ p53 axis may represent a new strategy for overcoming metastasis of CRC.

Background/Aims: Mesenchymal stem cells (MSCs) play an important role in regulating angiogenesis and immune balance. Abnormal proliferation and function of MSCs were reported at maternal fetal interface in patients with pre-eclampsia (PE). Micro-RNA-495 was known to be upregulated in the MSCs derived from patients with PE. However, it is not clear whether the up-regulated miR-495 is related to the pathogenesis of PE. Methods: We analyzed the expression of miR-495 in MSCs and umbilical cords derived from healthy pregnancies (NC) and PE, then we upregulated or downregulated the expression of miR-495 in MSCs derived from NC and tested the proliferation, apoptosis, migration, invasion, tube formation and senescence. Results: In the current study, we found that the expression of miR-495 was significantly increased in both umbilical cord tissues and MSCs in patients with severe PE. Overexpressing miR-495 arrested cell cycle in S phase and promoted cell apoptosis. The supernatants from miR-495-overexpressed-MSCs inhibited the migration of MSCs and HTR-8/SVneo, invasion of HTR-8/SVneo and tube formation of HUVEC, while si-miR-495 had the opposite effects. Furthermore, we analyzed the senescence related β-galactosidase activity and CD146 and found that miR-495 induced the senescence of MSCs. Molecular mechanism studies confirmed that Bmi-1 mediated these effects of miR-495 on MSCs. Conclusion: Taken together, our data demonstrated that miR-495 induced senescence of MSCs may be involved in the pathogenesis of PE.

MicroRNA 495 (miR-495) has been discovered to be involved in the metabolism and immune response in human body. The purpose of this study was to investigate the effect of miR-495 on macrophage M1/M2 polarization and insulin resistance in type 2 diabetes (T2D). A T2D mouse model was established by feeding C57BL/6 mice with a high-fat diet (HFD). The expressions of M1/M2 polarization markers and miR-495 in peritoneal macrophages were determined by qRT-PCR or Western blot. Mouse insulin tolerance test (ITT) and glucose tolerance test (GTT) were performed, and the targeted binding effect between miR-495, fat mass, and obesity-associated gene (FTO) was verified by double luciferase gene reporter assay. The body weight, blood glucose content, and miR-495 expression in macrophages of the HFD group were remarkably higher than those of the normal diet (ND) group. Besides, miR-495 induced the transformation of macrophages into M1-type pro-inflammatory macrophages and enhanced the insulin resistance of T2D mice. More importantly, FTO was proved to be a direct target gene of miR-495 and silencing FTO could induce the transformation of macrophages into M1-type pro-inflammatory macrophages. These results demonstrated that miR-495 could promote the transformation of macrophages into M1-type pro-inflammatory macrophages by inhibiting the expression of its target gene FTO, and aggravate the insulin resistance and adipose tissue inflammation in T2D mice, which provided a certain theoretical basis for the targeted treatment of T2D. [ABSTRACT FROM AUTHOR]

Choriocarcinoma is a rare gestational trophoblastic tumour. Propofol is one of the commonly used intravenous anaesthetic agents and exerts an anti-tumour role in multiple cancers. However, the effect of propofol on choriocarcinoma has not been well explored. We investigated the effect of propofol on JEG-3 cells. The effects of propofol on cell proliferation and metastasis were determined. The expression of miR-495 was suppressed while Twist1 overexpressed. Cell viability, migration and invasion were respectively measured using CCK-8 assay and transwell migration/invasion assay. The expression of miR-495, AKT1 and Twist1 was detected using qRT-PCR. Western blot analysis was performed to assess the expression levels of EMT- and proliferation-related factors, and TGF-β pathway proteins. Propofol treatment significantly inhibited cell proliferation, migration and invasion, and reduced the releasing of EMT. miR-495 was up-regulated by profocol, and miR-495 inhibition reversed the effect of propofol on cell proliferation and metastasis in JEG-3 cells. Furthermore, propofol decreased the expression of AKT1 and Twist1 through up-regulating miR-495, and Twist1 participated in the regulation of propofol on TGF-β signalling inactivation in JEG-3 cells. This study demonstrates that propofol suppresses cell proliferation and metastasis through up-regulation of miR-495 and down-regulation of Twist1 in JEG3 cells. Propofol inhibits proliferation and metastasis in JEG-3 cells; miR-495 is up-regulated by propofol in JEG-3 cells; miR-495 suppression blocks the effect of propofol in JEG-3 cells; Propofol decreases Twist1 expression through up-regulating miR-495; Propofol inhibits TGF-β signalling via regulation of Twist1. [ABSTRACT FROM AUTHOR]

Abstract Endometrial cancer (EC) is a huge threat to women's health. The aims of this study were to investigate the role of microRNA (miR)-495 in the proliferation and apoptosis of EC cells in vitro. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to detect the mRNA levels. In addition, dual-luciferase reporter assay was used to verified that PIK3R1 was a target of miR-495. After transfection with miR-495 mimics, Cell Counting Kit 8 (CCK-8) assay was performed to evaluate the cell viability of EC cells. The protein expression of PIK3R1, vascular endothelial growth factor (VEGF), Bcl-2, Bax, caspase 3 after transfection was analyzed using western blotting. Furthermore, cell apoptosis rate of EC cells was evaluated by flow cytometry. These results showed that miR-495 was significantly down-regulated in tumor tissues compared with the adjacent normal tissues, while PIK3R1 was up-regulated. The proliferation of the EC cells that were transfected with miR-495 mimics was markedly inhibited, and apoptosis was significantly promoted. In addition, downregulated expression of PIK3R1, Bcl-2, VEGF expression and upregulated expression of Bax and caspase 3 expression were observed after transfected with miR-495 mimic. Together these findings indicated that miR-495 acts as a tumor suppressor gene by directly targeting PIK3R1 at the post-transcriptional level in EC cells in vitro. [ABSTRACT FROM AUTHOR]

Pax3 plays an essential role in myogenesis. Previously, we found a tumor-signature chimeric fusion RNA, PAX3-FOXO1 also present during muscle differentiation, raising the possibility of its physiological role. Here we demonstrated that the fusion is needed transiently for muscle lineage commitment. Interestingly, the fusion ortholog was not found in seven mouse muscle differentiation/regeneration systems, nor in other stem cell differentiation systems of another three mammal species. We noticed that Pax3 is expressed at a much lower level in human stem cells, and during muscle differentiation than in other mammals. Given the fact that the fusion and the parental Pax3 share common downstream targets, we reasoned that forming the fusion may be a mechanism for human cells to escape certain microRNA regulation on Pax3. By sequence comparison, we identified 16 candidate microRNAs that may specifically target the human PAX3 3ʹUTR. We used a luciferase reporter assay, examined the microRNAs expression, and conducted mutagenesis on the reporters, as well as a CRISPR/Cas9 mediated editing on the endogenous allele. Finally, we identified miR-495 as a microRNA that specifically targets human PAX3. Examining several other fusion RNAs revealed that the human-specificity is not limited to PAX3-FOXO1. Based on these observations, we conclude that PAX3-FOXO1 fusion RNA is absent in mouse, or other mammals we tested, the fusion RNA is a mechanism to escape microRNA, miR-495 regulation in humans, and that it is not the only human-specific fusion RNA. [ABSTRACT FROM AUTHOR]

Background: Alterations in the expression level of miR-495 were recently observed in various tumours. Medulloblastoma is the most common malignant brain tumour in children. However, the clinical significance of miR-495 in medulloblastomas remains unclear. Methods: The expression levels of miR-495 was examined in 62 archival formalin-fixed paraffin-embedded (FFPE) medulloblastoma specimens using TaqMan Real-time Quantitative PCR arrays. Immunohistochemistry was used to determine the expression of Gfi1 in medulloblastoma tissues, and a luciferase reporter assay was carried out to confirm whether Gfi1 is a direct target of miR-495. Results: MiR-495 expression is repressed in medulloblastoma samples compared with normal cerebellum tissues. Furthermore, patients with a low level of miR-495 showed significantly poorer survival, as determined by the log-rank test (P = 0.033). The multivariate analysis results showed that the miR-495 expression levels were an independent predictor of overall survival in medulloblastoma patients (P = 0.027; hazard ratio = 0.267). Our study provides the first demonstration that miR-495 directly interacts with the Gfi1 3'UTR to regulate Gfi1 at a post-transcriptional level and that the expression level of miR-495 is inversely correlated with the Gfi1 protein level in medulloblastoma specimens. Conclusion: Our results suggest that miR-495 may be a prognostic predictor in medulloblastoma and that Gfi1 is a potential functional target of miR-495.

OBJECTIVE: Different effects of microRNA-495 (miR-495) on human cancers have been exhibited in recent years. However, the specific function of miR-495 remains uncertain in non-small-cell lung cancer (NSCLC). Thus, we aim to explore the role of miR-495 in NSCLC. PATIENTS AND METHODS: The expressions of miR-495 and transcription factor 4 (TCF4) were detected through quantitative Real-time polymerase chain reaction (qRT-PCR) assay. Western blot was used to measure the protein expression of relative genes. The relationship between miR-495 and TCF4 was testified by the dual-luciferase reporter gene assay. The function of miR-495 was investigated through cell counting kit-8 (CCK-8) assay and transwell assay. RESULTS: MiR-495 was downregulated in NSCLC tissues. Overexpression of miR-495 inhibited the migration, invasion and proliferation of NSCLC cells. Further, TCF4 was a direct target gene of miR-495. TCF4 was highly expressed in NSCLC tissues. In addition, miR-495 inhibited the progression of NSCLC through targeting TCF4. Furthermore, miR-495 inhibited EMT and Wnt/β-catenin pathway in NSCLC. CONCLUSIONS: MiR-495 inhibited the progression of NSCLC by targeting TCF4 and inactivating Wnt/β-catenin pathway. [ABSTRACT FROM AUTHOR]

High glucose (HG)-induced apoptosis of retinal ganglion cells (RGCs) contributes to the pathogenesis of diabetic retinopathy, which is one of the most common and severe complications of diabetes mellitus. Accumulating evidence has documented that microRNAs (miRNAs) play an important role in the pathogenesis of diabetic retinopathy. However, the role of miRNAs in regulating HG-induced apoptosis of RGCs remains largely unknown. Various studies have suggested that miR-495 is an important regulator of cell apoptosis and survival. In this study, we aimed to investigate whether miR-495 is involved in regulating HG-induced apoptosis of RGCs and reveal its possible relevance in diabetic retinopathy. We found that miR-495 was significantly upregulated in HG-treated RGCs. Downregulation of miR-495 protected RGCs against HG-induced apoptosis, whereas overexpression of miR-495 had the opposite effect. Notably, Notch1 was identified as a target gene of miR-495, as miR-495 negatively regulated Notch1 expression and the Notch signaling pathway. Moreover, downregulation of miR-495 inhibited PTEN expression while promoting Akt activation. However, knockdown of Notch1 significantly abolished the protective effect of miR-495 inhibition against HG-induced apoptosis. Overall, our study suggests that downregulation of miR-495 alleviates HG-induced apoptosis of RGCs by targeting Notch1 to regulate PTEN/Akt signaling, which provides novel insights into understanding the pathogenesis of HG-induced apoptosis of RGCs. [ABSTRACT FROM AUTHOR]

Background Both LncRNA UCA1 and miR-495 are crucial gene regulators in various disorders. This study aims to investigate their role in epilepsy and seizure-induced brain injury. Methods In this research, rat model of epilepsy was established by pilocarpine induction. The RNA and protein expression in hippocampal tissues and neurons were determined by qRT-PCR and western blot, respectively. The hippocampal neurons were isolated from hippocampal tissues, and treated with magnesium-free (MGF) physiological solution for epileptiform activity induction. The endogenous expression of related genes was modulated by recombinant plasmids and cell transfection. Flow cytometry was used to analyze the cell apoptosis. Dual luciferase reporter assay was performed to determine the interaction between miR-495 and Nrf2 in HEK-293 cells. Results The lncRNA UCA1 and Nrf2 were down-regulated in epileptiform hippocampal tissues and neurons, while the miR-495 was up-regulated. Over-expression of UCA1 inhibited apoptosis of hippocampal neurons by suppressing miR-495. MiR-495 negatively regulated Nrf2. UCA1 inhibited apoptosis of hippocampal neurons through miR-495/Nrf2-ARE pathway. UCA1 suppressed pilocarpine-induced epilepsy in rat. Conclusion LncRNA UCA1 suppressed pilocarpine-induced epilepsy by inhibiting apoptosis of hippocampal neurons through miR-495/Nrf2-ARE pathway, and thereby inhibiting brain injury induced by seizure. [ABSTRACT FROM AUTHOR]

Background: Amounting evidence indicate that miRNAs play an important role in the development of various cancers. MiR-495 is a potential tumor suppressor in cancers, however its role in melanoma is still elusive. The study aimed to investigate the role of miR-495 and the underlying mechanisms in melanoma cells. Methods: The levels of miR-495 in melanoma tissues and cell lines were measured by quantitative real-time polymerase chain reaction. Mimics of miR-495 was transfected into human melanoma cells A375 and MeWo. Cell viability of miR-495-transfected cells was assayed by MTT assay. Cell migration and invasion of miR-495 transfected cells were measured by wound healing assay and transwell assay, respectively. Nucleosome enzyme-linked immunosorbent assay was performed to measure the apoptosis induced by overexpression of miR-495. Luciferase reporter assays were performed to verify the interaction between miR-495 and its target PBX3. Results: It was found that the expression levels of miR-495 were down-regulated in melanoma tissues and cells. Moreover, overexpression of miR-495 inhibited melanoma cell proliferation, migration and invasion in vitro. PBX3 was identified as a target for inhibition by miR-495 and was confirmed by luciferase assay, quantitative real-time polymerase chain reaction and western blot. We also indicated that silencing of PBX3 also repressed melanoma cell proliferation, migration and invasion in vitro. Conclusion: In summary, our findings demonstrated that miR-495 functions as a tumor suppressor in human melanoma via directly targeting PBX3. [ABSTRACT FROM AUTHOR]

Background/Aims: More and more reports have shown that the dysregulation of miRNAs can contribute to the progression and metastasis of human cancers. Many studies have shown that the down-regulation of the miR-495 level occurs in a variety of cancers, including colorectal cancer (CRC). However, the precise molecular mechanisms of miR-495 in CRC have not been well clarified. In the current study, we investigated the biological functions and molecular mechanisms of miR-495 in CRC cell lines. Methods: qRT-PCR was used to determine the level of miR-495 in CRC cell lines and tissues. A miR-495 mimic and inhibitor were transfected into CRC cells, and the effects of miR-495 on the invasion and EMT were explored by qRT-PCR as well as transwell and Western blot assays. Meanwhile, luciferase assays were performed to validate Annexin A3 as a miR-495 target in CRC cells. Results: In our study, we found that miR-495 is down-regulated in CRC tissues and cell lines. Moreover, the low level of miR-495 was associated with increased expression of Annexin A3 in CRC tissues and cell lines. The invasion and EMT of CRC cells were suppressed by the overexpression of miR-495. However, the down-regulation of miR-495 promoted the invasion and EMT of CRC cells. Bioinformatics analysis predicted that Annexin A3 was a potential target gene of miR-495. Next, the luciferase reporter assay confirmed that miR-495 could directly target Annexin A3. Consistent with the effect of miR-495, the down-regulation of Annexin A3 by siRNA inhibited the invasion and EMT of CRC cells through the up-regulation of p53. The introduction of Annexin A3 in CRC cells partially blocked the effects of the miR-495 mimic. Conclusion: The introduction of miR-495 directly targeted Annexin A3 to inhibit the invasion and EMT of CRC cells by up-regulating p53, and the down-regulation of Annexin A3 was essential for inhibiting the invasion and EMT of CRC cells by overexpressing miR-495. Overall, the re-activation of the miR-495/Annexin A3/ p53 axis may represent a new strategy for overcoming metastasis of CRC. [ABSTRACT FROM AUTHOR]

microRNAs (miRNAs) have been reported to play crucial roles in malignant tumor progression, including cell development, proliferation, and progression. Increasing evidence suggests that miR-495 functions as either an oncogene or tumor suppressor in several tumor types. However, its biological role in the development of colorectal cancer (CRC) still remains unclear. In this study, we found that miR-495 expression level was remarkably down-regulated in CRC tissues samples and cell lines when compared to adjacent normal tissues and cell line by using qRT-PCR detection. Ectopic expression of miR-495 by mimic transfection significantly suppressed CRC cell proliferation and colony formation, inhibited migration and invasion, and induced apoptosis according to CCK-8, colony formation, transwell, and flow cytometry assays. Furthermore, bioinformatics and luciferase reporter assays verified that family with sequence similarity 83, member D (FAM83D) was a direct target of miR-495 in CRC cells, and FAM83D was confirmed to be upregulated in human CRC tissues and reversely correlated with miR-495 expression. In addition, rescue experiments revealed that restoration of FAM84D could partially abrogate the inhibitory effect of miR-495 overexpression on CRC cell proliferation and metastasis. Further mechanic investigations also verified that the PTEN/P13K/AKT/mTOR pathway was involved in the miR-495/FAM83D-mediated CRC cell progression. Our findings identified that miR-495 acted as a critical tumor suppressor by directly targeting FAM83D during CRC development and therefore represented as a potential biomarker for CRC therapy. [ABSTRACT FROM AUTHOR]

MDR1 is highly expressed in MDR A2780DX5 ovarian cancer cells, MDR SGC7901R gastric cancer cells and recurrent tumours. It pumps cytoplasmic agents out of cells, leading to decreased drug accumulation in cells and making cancer cells susceptible to multidrug resistance. Here, we identified that miR-495 was predicted to target ABCB1, which encodes protein MDR1. To reduce the drug efflux and reverse MDR in cancer cells, we overexpressed a miR-495 mimic in SGC7901R and A2780DX cells and in transplanted MDR ovarian tumours in vivo. The results indicated that the expression of MDR1 in the above cells or tumours was suppressed and that subsequently the drug accumulation in the MDR cells was decreased, cell death was increased, and tumour growth was inhibited after treatment with taxol-doxorubicin, demonstrating increased drug sensitivity. This study suggests that pre-treatment with miR-495 before chemotherapy could improve the curative effect on MDR1-based MDR cancer. [ABSTRACT FROM AUTHOR]

Background/Aims: Mesenchymal stem cells (MSCs) play an important role in regulating angiogenesis and immune balance. Abnormal proliferation and function of MSCs were reported at maternal fetal interface in patients with pre-eclampsia (PE). Micro-RNA-495 was known to be upregulated in the MSCs derived from patients with PE. However, it is not clear whether the up-regulated miR-495 is related to the pathogenesis of PE. Methods: We analyzed the expression of miR-495 in MSCs and umbilical cords derived from healthy pregnancies (NC) and PE, then we upregulated or downregulated the expression of miR-495 in MSCs derived from NC and tested the proliferation, apoptosis, migration, invasion, tube formation and senescence. Results: In the current study, we found that the expression of miR- 495 was significantly increased in both umbilical cord tissues and MSCs in patients with severe PE. Overexpressing miR-495 arrested cell cycle in S phase and promoted cell apoptosis. The supernatants from miR-495-overexpressed-MSCs inhibited the migration of MSCs and HTR- 8/SVneo, invasion of HTR-8/SVneo and tube formation of HUVEC, while si-miR-495 had the opposite effects. Furthermore, we analyzed the senescence related β-galactosidase activity and CD146 and found that miR-495 induced the senescence of MSCs. Molecular mechanism studies confirmed that Bmi-1 mediated these effects of miR-495 on MSCs. Conclusion: Taken together, our data demonstrated that miR-495 induced senescence of MSCs may be involved in the pathogenesis of PE. [ABSTRACT FROM AUTHOR]

Long non-coding RNAs have recently emerged as important regulators in the pathogenesis and progression of cancers. The long non-coding RNA urothelial carcinoma-associated 1 is reportedly upregulated and functions as an oncogene in some tumors. However, the role of urothelial carcinoma-associated 1 in renal cell carcinoma is not well elucidated so far. In this study, we found that urothelial carcinoma-associated 1 was overexpressed in renal cell carcinoma tissues compared with the adjacent normal tissues, and higher urothelial carcinoma-associated 1 expression levels were positively associated with advanced tumor stage and poor survival time in renal cell carcinoma patients. Further studies showed that knockdown of urothelial carcinoma-associated 1 suppressed renal cell carcinoma cell proliferation and S-phase cell number in vitro. Moreover, urothelial carcinoma-associated 1 was found to be associated with enhancer of zeste homolog 2, which suppressed p21 expression through histone methylation (H3K27me3) on p21 promoter. We also showed that knockdown of urothelial carcinoma-associated 1 increased the p21 protein expression through regulating enhancer of zeste homolog 2. In addition, bioinformatics analysis and dual-luciferase reporter assays confirmed that miR-495 was a target of urothelial carcinoma-associated 1 in renal cell carcinoma, and urothelial carcinoma- associated 1 promoted cell proliferation by negatively regulating miR-495. These findings illuminated that urothelial carcinoma-associated 1 promoted renal cell carcinoma progression through enhancer of zeste homolog 2 and interacted with miR-495. Overall, overexpression of urothelial carcinoma-associated 1 functions as an oncogene in renal cell carcinoma that may offer a novel therapeutic target for renal cell carcinoma patients. [ABSTRACT FROM AUTHOR]

Studies have shown that TSPAN12 serves as an oncogenic gene or tumor suppressor depending on the types of cancer. However, its role in the drug resistance of small cell lung cancer (SCLC) is still unknown. In this study, we investigated whether TSPAN12 regulates chemoresistance, proliferation and tumor growth of SCLC under the regulation of miR-495 using two drug resistant cell lines. The results showed that down-regulation of TSPAN12 inhibited cells chemoresistance, proliferation and tumor growth. Besides, TSPAN12 elevation in SCLC specimens correlated with poor pathologic stage and shorter survival time. In addition, the dual luciferase assay indicated that TSPAN12 was one of the directly targeted genes of miR-495. Our study revealed that TSPAN12 promoted chemoresistance of SCLC under the regulation of miR-495. TSPAN12 depletion is a promising strategy for inhibiting the chemoresistance in SCLC. [ABSTRACT FROM AUTHOR]

Accumulating evidence has linked deregulation of microRNA-495 (miR-495) to tumorigenesis; however, its function in tumor progression is controversial. This work was undertaken to explore the expression and biological roles of miR-495 in bladder cancer. The expression of miR-495 was examined in 67 pairs of bladder cancer and adjacent normal bladder tissues. The roles of miR-495 in bladder cancer cell proliferation and invasion in vitro and tumorigenesis in vivo were determined. Direct target gene(s) mediating the activity of miR-495 in bladder cancer cells was identified. It was found that miR-495 was expressed at greater levels in bladder tissues and cell lines. High expression of miR-495 was significantly associated with larger tumor size, advanced TNM stage, and lymph node metastasis. Overexpression of miR-495 significantly promoted bladder cancer cell proliferation and invasion, whereas inhibition of miR-495 suppressed cell proliferation and invasion. PTEN, a well-defined tumor suppressor was identified to be a target gene of miR-495. A significant inverse correlation between miR-495 and PTEN expression was noted in bladder cancer tissues ( r = −0.3094, P = 0.0125). Overexpression of miR-495 led to reduction of PTEN expression in bladder cancer cells. Rescue experiments showed that enforced expression of PTEN impaired miR-495-mediated bladder cancer proliferation and invasion. In vivo mouse studies demonstrated that overexpression of miR-495 accelerated the growth of subcutaneous bladder cancer xenografts, which was associated with downregulation of PTEN. Overall, these findings indicate that miR-495 upregulation contributes to bladder cancer cell growth, invasion, and tumorigenesis by targeting PTEN and offer a potential therapeutic target for bladder cancer. [ABSTRACT FROM AUTHOR]