Background:Glioblastoma remains a highly invasive primary brain malignancy with an undesirable prognosis.Growing evidence has shed light on the importance of microRNAs(miRs),as small non-coding RNAs,in tumor developme...Background:Glioblastoma remains a highly invasive primary brain malignancy with an undesirable prognosis.Growing evidence has shed light on the importance of microRNAs(miRs),as small non-coding RNAs,in tumor development and progression.The present study leverages the in-silico and in-vitro techniques to investigate the significance of hsa-miR-181a-5p and the underlying hsa-miR-181a-5p-meidated signaling pathway in glioblastoma development.Methods:Bioinformatic studies were performed on GSE158284,GSE108474(REMBRANDT study),TCGA-GTEx,CCLE,GeneMANIA,Reactome,WikiPathways,KEGG,miRDB,and microT-CDS to identify the significance of hsa-miR-181a-5p and its underlying target.Afterward,the U373 cell line was selected and transfected with hsa-miR-181a-5p mimics,and the cell viability,clonogenicity,migration,mRNA expression,apoptosis,and cell cycle were studied using the MTT assay,colony formation test,migration assay,qRT-PCR,andflow cytometry respectively.Results:hsa-miR-181a-5p expression is decreased in glioblastoma samples.The in-silico results have shown that hsa-miR-181a-5p could regulate the MAPK pathway by targeting AKT3.The experimental assays have shown that hsa-miR-181a-5p decreases the migration of glioblastoma cells,arrests the cell cycle,and increases the apoptosis rate.Besides downregulating MMP9 and upregulating BAX,hsa-miR-181a-5p downregulates MET,MAP2K1,MAPK1,MAPK3,and AKT3 expression in U373 cells.The in-vitro results were consistent with in-silico results regarding the regulatory effect of hsa-miR-181a-5p on the MAPK pathway,leading to tumor suppression in glioblastoma.Conclusions:hsa-miR-181a-5p inhibits glioblastoma development partially by regulating the signaling factors of the MAPK pathway.展开更多
MicroRNAs(miRNAs)have been demonstrated to control chicken skeletal muscle growth,however,the potential function of the miR-181-5p family in chicken myogenesis remains largely unknown.Here,our study identified the two...MicroRNAs(miRNAs)have been demonstrated to control chicken skeletal muscle growth,however,the potential function of the miR-181-5p family in chicken myogenesis remains largely unknown.Here,our study identified the two chicken(Gallus gallus;Gga)miR-181-5p family members widely expressed in various tissues,specifically miR-181a-5p and miR-181b-5p.Besides,the breast muscles of fast-growing broilers expressed higher levels of miR-181a-5p and miR-181b-5p than those of slow-growing layers.Functionally,miR-181a-5p and miR-181b-5p both promote the expression level of myogenic factors including myogenin(MyoG),myogenic differentiation 1(MyoD1),and myosin heavy chain(MyHC),meanwhile accelerating the myotube formation of skeletal muscle satellite cells(SMSCs).Mechanistically,miR-181a-5p and miR-181b-5p directly bind to the 3′untranslated region(UTR)of the transforming growth factor beta receptor 1(TGFBR1)mRNA,further reducing the expression of TGFBR1.TGFBR1 is a key Transforming growth factor beta(TGF-β)signaling transduction receptor and had a negative function in muscle cell differentiation.Furthermore,knockdown of TGFBR1 facilitated the expression of chicken myogenic factors,boosted myotube formation,and decreased the SMAD family member 2/3(SMAD2/3)phosphorylation in chicken SMSCs.SMAD2/3 are downstream of TGF-βsignaling,and miR-181a-5p and miR-181b-5p could reduce the expression of TGFBR1 to further diminish the SMAD2/3 phosphorylation.Our findings revealed that the miR-181-5p family targets TGFBR1 to break the TGF-βsignaling transduction,which resulted in promoting chicken skeletal muscle development.展开更多
目的研究miR-181a、半乳凝素-3(galectin-3,GAL3)在急性缺血性脑卒中(acute ischemic stroke,AIS)病情及预后评估中的临床价值。方法选取2019年1月至2021年1月诊治的82例AIS患者(AIS组)和40例同期无缺血性脑卒中的颅脑血管疾病患者(对照...目的研究miR-181a、半乳凝素-3(galectin-3,GAL3)在急性缺血性脑卒中(acute ischemic stroke,AIS)病情及预后评估中的临床价值。方法选取2019年1月至2021年1月诊治的82例AIS患者(AIS组)和40例同期无缺血性脑卒中的颅脑血管疾病患者(对照组)为研究对象,比较两组患者的血清miR-181a、GAL3、美国国立卫生研究院卒中量表(National Institute of Health stroke scale,NIHSS)、改良Barthel指数评定量表(modified Barthel index,MBI)及Fugl-Meyer肢体运动功能(Fugl-Meyer assessment of gait,FMA)评分的差异。Pearson相关性分析miR-181a、GAL3与神经功能指标(NIHSS、MBI、FMA评分)的相关性。采用受试者操作特征曲线(receiver operating characteristic curve,ROC曲线)分析miR-181a、GAL3预测AIS患者1年预后不良的效能。Logistic回归分析AIS患者1年死亡的危险因素。结果与对照组比较,AIS组患者的miR-181a、GAL3、NIHSS评分显著增高(P<0.05);MBI及FMA评分均显著降低(P<0.05)。AIS组患者的miR-181a、GAL3与NIHSS评分呈正相关(r>0,P<0.05);与MBI评分、FMA评分呈负相关(r<0,P<0.05)。miR-181a、GAL3预测AIS组患者1年预后不良的敏感度、特异性、曲线下面积(areaunder the curve,AUC)均高于NIHSS评分、MBI评分、FMA评分;miR-181a联合GAL3预测AIS组患者1年预后不良的敏感度、特异性、AUC均高于miR-181a、GAL3、NIHSS评分、MBI评分、FMA评分。miR-181a≥1.3、GAL3≥12.3ng/ml、NIHSS评分≥17.4分、MBI评分<67.6分、FMA评分<84.2分为AIS患者1年死亡的独立危险因素(P<0.05)。结论AIS患者血清miR-181a及GAL3水平明显增高,在AIS病情及预后评估中具有一定的临床价值。展开更多
文摘Background:Glioblastoma remains a highly invasive primary brain malignancy with an undesirable prognosis.Growing evidence has shed light on the importance of microRNAs(miRs),as small non-coding RNAs,in tumor development and progression.The present study leverages the in-silico and in-vitro techniques to investigate the significance of hsa-miR-181a-5p and the underlying hsa-miR-181a-5p-meidated signaling pathway in glioblastoma development.Methods:Bioinformatic studies were performed on GSE158284,GSE108474(REMBRANDT study),TCGA-GTEx,CCLE,GeneMANIA,Reactome,WikiPathways,KEGG,miRDB,and microT-CDS to identify the significance of hsa-miR-181a-5p and its underlying target.Afterward,the U373 cell line was selected and transfected with hsa-miR-181a-5p mimics,and the cell viability,clonogenicity,migration,mRNA expression,apoptosis,and cell cycle were studied using the MTT assay,colony formation test,migration assay,qRT-PCR,andflow cytometry respectively.Results:hsa-miR-181a-5p expression is decreased in glioblastoma samples.The in-silico results have shown that hsa-miR-181a-5p could regulate the MAPK pathway by targeting AKT3.The experimental assays have shown that hsa-miR-181a-5p decreases the migration of glioblastoma cells,arrests the cell cycle,and increases the apoptosis rate.Besides downregulating MMP9 and upregulating BAX,hsa-miR-181a-5p downregulates MET,MAP2K1,MAPK1,MAPK3,and AKT3 expression in U373 cells.The in-vitro results were consistent with in-silico results regarding the regulatory effect of hsa-miR-181a-5p on the MAPK pathway,leading to tumor suppression in glioblastoma.Conclusions:hsa-miR-181a-5p inhibits glioblastoma development partially by regulating the signaling factors of the MAPK pathway.
基金supported by the National Key Research and Development Program of China(2022YFF10002020)Sichuan Science and Technology Program,China(2021YFYZ0007 and 2021YFYZ0031).
文摘MicroRNAs(miRNAs)have been demonstrated to control chicken skeletal muscle growth,however,the potential function of the miR-181-5p family in chicken myogenesis remains largely unknown.Here,our study identified the two chicken(Gallus gallus;Gga)miR-181-5p family members widely expressed in various tissues,specifically miR-181a-5p and miR-181b-5p.Besides,the breast muscles of fast-growing broilers expressed higher levels of miR-181a-5p and miR-181b-5p than those of slow-growing layers.Functionally,miR-181a-5p and miR-181b-5p both promote the expression level of myogenic factors including myogenin(MyoG),myogenic differentiation 1(MyoD1),and myosin heavy chain(MyHC),meanwhile accelerating the myotube formation of skeletal muscle satellite cells(SMSCs).Mechanistically,miR-181a-5p and miR-181b-5p directly bind to the 3′untranslated region(UTR)of the transforming growth factor beta receptor 1(TGFBR1)mRNA,further reducing the expression of TGFBR1.TGFBR1 is a key Transforming growth factor beta(TGF-β)signaling transduction receptor and had a negative function in muscle cell differentiation.Furthermore,knockdown of TGFBR1 facilitated the expression of chicken myogenic factors,boosted myotube formation,and decreased the SMAD family member 2/3(SMAD2/3)phosphorylation in chicken SMSCs.SMAD2/3 are downstream of TGF-βsignaling,and miR-181a-5p and miR-181b-5p could reduce the expression of TGFBR1 to further diminish the SMAD2/3 phosphorylation.Our findings revealed that the miR-181-5p family targets TGFBR1 to break the TGF-βsignaling transduction,which resulted in promoting chicken skeletal muscle development.
文摘目的研究miR-181a、半乳凝素-3(galectin-3,GAL3)在急性缺血性脑卒中(acute ischemic stroke,AIS)病情及预后评估中的临床价值。方法选取2019年1月至2021年1月诊治的82例AIS患者(AIS组)和40例同期无缺血性脑卒中的颅脑血管疾病患者(对照组)为研究对象,比较两组患者的血清miR-181a、GAL3、美国国立卫生研究院卒中量表(National Institute of Health stroke scale,NIHSS)、改良Barthel指数评定量表(modified Barthel index,MBI)及Fugl-Meyer肢体运动功能(Fugl-Meyer assessment of gait,FMA)评分的差异。Pearson相关性分析miR-181a、GAL3与神经功能指标(NIHSS、MBI、FMA评分)的相关性。采用受试者操作特征曲线(receiver operating characteristic curve,ROC曲线)分析miR-181a、GAL3预测AIS患者1年预后不良的效能。Logistic回归分析AIS患者1年死亡的危险因素。结果与对照组比较,AIS组患者的miR-181a、GAL3、NIHSS评分显著增高(P<0.05);MBI及FMA评分均显著降低(P<0.05)。AIS组患者的miR-181a、GAL3与NIHSS评分呈正相关(r>0,P<0.05);与MBI评分、FMA评分呈负相关(r<0,P<0.05)。miR-181a、GAL3预测AIS组患者1年预后不良的敏感度、特异性、曲线下面积(areaunder the curve,AUC)均高于NIHSS评分、MBI评分、FMA评分;miR-181a联合GAL3预测AIS组患者1年预后不良的敏感度、特异性、AUC均高于miR-181a、GAL3、NIHSS评分、MBI评分、FMA评分。miR-181a≥1.3、GAL3≥12.3ng/ml、NIHSS评分≥17.4分、MBI评分<67.6分、FMA评分<84.2分为AIS患者1年死亡的独立危险因素(P<0.05)。结论AIS患者血清miR-181a及GAL3水平明显增高,在AIS病情及预后评估中具有一定的临床价值。
