MicroRNA-323 regulates ischemia/reperfusion injury-induced neuronal cell death by targeting BRI3

Purpose: MicroRNA-323 (miR-323) has been reported to be upregulated in Ischemia/Reperfusion (I/R) injury-treated neuronal cell. However, the effect and underlying mechanism of miR-323 in I/R-induced neuronal cell death remains poorly understood. The current study was aim to investigate the role and molecular basis of miR-323 in I/R-induced neuronal cell. Methods: An oxygen-glucose deprivation (OGD) model of hippocampal neuron I/R was produced in vitro. Cell apoptosis, cell survival, and the expression of miR-323 were determined after 6 h, 12 h and 24 h after OGD treatment. The up- or down-regulation of miR-323 was performed by miR-323 mimics or anti-miR-323, respectively. Results: OGD induced apoptosis and suppressed survival in rat hippocampal neurons. And the expression levels of miR-323 were increased after OGD treatment. Furthermore, the up-regulation of miR-323 promoted apoptosis and suppressed survival, whereas the inhibition of miR-323 suppressed apoptosis and enhanced survival in OGD-treated neurons. Moreover, miR-323 could directly bind to BRI3 3’-UTR. Notably, the knockdown of BRI3 by BRI3 siRNA apparently abrogated cell survival and induced cell apoptosis in rat neurons. Conclusion: This study indicated that miR-323 might regulate ischemia/reperfusion-induced rat neuronal cell death via targeting BRI3.     

MicroRNA4293基因多态性与女性肝癌易感性相关

目的研究microRNA-4293的SNP(rs12220909)与原发性肝癌易感性的关系。方法收集人体血液样本1788份,样本取自健康对照者884名,原发性肝癌患者904例。抽提血液样本中基因组DNA,应用MassARRAY法对microRNA-4293的SNP(rs12220909)位点进行基因分型;应用二元逻辑回归法分析microRNA-4293的SNP(rs12220909)位点与肝癌易感性之间的关系。结果女性肝癌患者microRNA-4293的SNP(rs12220909)中CC基因型较健康对照组显著增加(P=0.04),在HBV相关肝癌中显著性进一步增加(P=0.02),提示CC基因可能增加女性肝癌尤其是HBV相关肝癌的易感性。而在男性患者中,microRNA-4293的SNP(rs12220909)与肝癌易感性并不相关(P=0.33)。结论microRNA-4293的SNP(rs12220909)与女性肝癌易感性相关。     

miR-21反义寡核苷酸增强地西他滨体外抗白血病效应

目的:研究miR-21反义寡核苷酸(anti-miR-21 oligonucleotide,AMO)对地西他滨(decitabine,DCA)抗白血病效应的影响及可能机制。方法:将AMO和无义寡核苷酸(scramble oligonucleotide,SCR)通过脂质体转染导入HL-60细胞,实时荧光定量PCR(real-time PCR)验证转染效率,再分别与DCA 0.5、2.0和4.0μmol/L作用48 h。Real-time PCR分别检测人周期节律蛋白3(h Per3)mRNA表达,Annexin V/PI法检测凋亡,流式细胞术检测CD117和CD11b平均荧光强度(MFI)。结果:AMO转染组miR-21表达(0.35±0.07)低于空白组(0.71±0.07)和SCR转染组(0.66±0.05),差异有统计学意义(P0.05)。AMO转染组的HL-60细胞DCA的IC50低于空白组和SCR转染组(P0.01)。同一浓度下,AMO组的早期凋亡率、CD11b的MFI和h Per3 mRNA均高于同一浓度药物作用的空白组和SCR组,CD117 MFI均低于同一浓度药物作用的空白组和SCR组,差异均具有统计学意义(P0.01)。结论:AMO能显著促进DCA体外抗白血病效应,其机制可能与其协助激活h Per3的表达有关。     

miR-486-5p通过抑制端粒酶活性促进人骨髓间充质干细胞衰老

目的:研究年龄相关microRNA-486-5p(miR-486-5p)对人骨髓间充质干细胞(h MSCs)衰老的调控作用。方法:通过microRNA芯片和real-time PCR检测供体年龄对h MSCs中miR-486-5p表达的影响;通过转染miR-486-5p模拟物或抑制物,过表达miR-486-5p或抑制其表达;用β-半乳糖苷酶染色检测miR-486-5p对h MSCs衰老的影响;通过siRNA研究沉默信息调节因子1(SIRT1)对h MSCs端粒酶逆转录酶(TERT)、端粒酶活性及衰老的影响。结果:随供体年龄增加,h MSCs中miR-486-5p的表达增加。过表达miR-486-5p可促进h MSCs衰老。相反,抑制miR-486-5p的表达可减少h MSCs的衰老。SIRT1及TERT随供体年龄增加而表达下降,直接抑制SIRT1的表达可减少TERT表达,抑制端粒酶活性,促进细胞衰老。同时抑制miR-486-5p和SIRT1的表达,使miR-486-5p失去对h MSCs端粒酶活性及衰老的调控作用。结论:miR-486-5p通过抑制SIRT1,减少h MSCs端粒酶活性,促进h MSCs衰老。     

miR-21在周围神经损伤时调控雪旺细胞凋亡

目的:探讨周围神经损伤时,微小RNA-21(microRNA-21,miR-21)与雪旺细胞(SC)凋亡的关系及相关分子机制。方法:采用实时荧光定量PCR(real-time PCR)检测动物模型中miR-21以及人第10号染色体缺失的磷酸酶及张力蛋白同源基因(phosphatase and tensin homologue deleted on chromosome 10,PTEN)的表达情况;通过将miR-21类似物(miR-21-mimic)、miR-21抑制物(miR-21-inhibitor)和阴性对照miRNA(negative control miRNA,NC-miRNA)转染入RSC96细胞中,构建出过表达miR-21的雪旺细胞株(MI-SC)、抑制miR-21表达的雪旺细胞株(IN-SC)及表达对照miRNA的雪旺细胞株(NC-SC);采用流式细胞仪检测3组细胞的凋亡情况;real-time PCR检测3组细胞中miR-21以及PTEN的表达情况;Western blot检测3组细胞中PTEN及凋亡相关蛋白激活型caspase-3(cleaved caspase-3)的表达情况。结果:神经损伤组miR-21的表达量与对照组相比明显升高,神经损伤组PTEN mRNA的表达水平与对照组相比明显降低;与NC-SC相比,上调miR-21组细胞的凋亡比例减少,PTEN mRNA及蛋白的表达水平降低,cleaved caspase-3的表达水平降低,下调miR-21组细胞的凋亡比例增加,PTEN mRNA及蛋白的表达水平升高,cleaved caspase-3的表达水平升高(P0.05)。结论:miR-21可能通过下调PTEN的表达抑制雪旺细胞的凋亡,从而可能在周围神经的损伤修复中发挥作用。     

miR-1-3p在食蟹猴心肌损伤中表达水平及其与CK-MB、cTnI和NT-proBNP的相关性

目的探讨循环miR-1-3p在多柔比星诱导食蟹猴心肌损伤模型中的表达水平和诊断价值。方法选取3~5岁普通级雄性食蟹猴17只,随机分为对照组(7只)和实验组(10只),静脉注射盐酸多柔比星注射液(2.5 mg/kg)4周,收集外周血浆标本和心脏组织标本,应用液相芯片技术测定肌酸激酶同工酶(CK-MB)、肌钙蛋白I(cTnI)和NT-proBNP表达水平,且应用实时定量PCR技术测定miR-1-3p表达的水平,并进行相关性分析。结果成功建立药物致食蟹猴心肌损伤模型;两组CK-MB、cTnI和NT-proBNP水平比较差异有统计学意义(P<0.05);实验组miR-1-3p的表达水平在给药4周后较给药前和对照组相比明显升高(P<0.05);miR-1-3p的表达水平与CK-MB、cTnI和NT-proBNP改变呈现了相同的升高趋势,且二者之间具有明显相关性。结论miR-1-3p可作为药物引起心肌损伤的生物学诊断标志物之一。