微小RNA-548z与CD147基因3′UTR结合对急性心肌梗死患者CD147表达的影响
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(江苏大学附属医院心内科,江苏省镇江市 212001)

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梁潇,硕士研究生,主要从事动脉粥样硬化研究。

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国家自然科学基金资助项目(81170279、81370409),江苏省六大人才高峰项目(WS-074),江苏省333高层次人才项目(BRA2014162),江苏省科教兴卫工程(LJ201116),镇江市心血管病重点实验室项目(SS2012002)


The Effect on the Expression of CD147 in Acute Myocardial Infarction Patients as MicroRNA-548z Binding to the CD147 Gene 3′ Untranslated Region
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Department of Cardiology,Affiliated Hospital of Jiangsu University,Zhenjiang,Jiangsu 212001,China)

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    摘要:

    目的 探讨微小RNA-548z(microRNA-548z, miR-548z)与CD147基因3′非翻译区单核苷酸多态性位点rs8259 T/A结合对急性心肌梗死(AMI)患者CD147表达的影响。方法 以30例AMI患者及13例正常人作为研究对象,应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术检测CD147基因rs8259位点的基因型。外周血单个核细胞(PBMC)中miR-548z及CD147 mRNA的表达采用荧光定量聚合酶链反应(qPCR)法检测,CD147蛋白表达应用Western Blot法。双荧光素酶报告基因验证miR-548z与rs8259 T/A的相互作用。 结果 AMI患者AA型和TT型PBMCs中miR-548z的表达水平无差异,并与正常对照组无统计学意义(P>0.05);AMI患者AA型和TT型PBMCs中CD147 mRNA的表达水平无差异(P>0.05);而AMI患者AA型组CD147蛋白的表达水平明显高于TT型组;双荧光素酶报告基因结果显示,mimic miR-548z能够抑制携带CD147基因rs8259位点T等位基因载体的荧光素酶活性,呈剂量依赖性。结论 miR-548z能与AMI患者CD147基因3′UTR rs8259的T等位基因结合,从而调控CD147蛋白的表达。

    Abstract:

    Aim To investigate the effect on the expression of CD147 in acute myocardial infarction (AMI) patients as microRNA-548z (miR-548z) binding to the single nucleotide polymorphism (SNP) sites rs8259 T / A at CD147 gene 3′untranslated region (UTR). Methods Samples for the study were from 30 patients with AMI and 13 healthy people. Genotype of rs8259 sites on CD147 gene was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Quantitative polymerase chain reaction (qPCR) was used to detect the relative expression levels of miR-548z and CD147 mRNA in peripheral blood mononuclear cells (PBMC). The expression of CD147 protein in PBMCs was detected by Western Blot. The interaction between miR-548z and rs8259 different alleles was proved by luciferase reporter gene assay. Results The expression of miR-548z in AA and TT AMI patients, as well as in normal people appeared no significant difference (P>0.05). There was no significant difference between AA and TT patients with AMI in the expression levels of CD147 mRNA in PBMCs (P>0.05). The level of CD147 protein in AMI patients with AA genotype was significantly higher than the TT genotype group. The luciferase reporter gene assay showed that mimic miR-548z could does-dependly reduce the relative luciferase activity of constructs carrying the T allele. Conclusion MiR-548z could bind to rs8259 bearing T alleles on the 3′UTR of CD147 gene and negatively regulate the expression of CD147 protein in AMI patients.

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梁潇,严金川,王翠平,王中群,刘培晶,袁伟.微小RNA-548z与CD147基因3′UTR结合对急性心肌梗死患者CD147表达的影响[J].中国动脉硬化杂志,2016,24(1):23~28.

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  • 收稿日期:2015-01-30
  • 最后修改日期:2015-03-20
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  • 在线发布日期: 2018-11-19