miR-372调控血管生成因子AGGF1的表达并抑制血管生成
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国家自然科学基金(81402550);江苏省自然科学基金(BK20140906);江苏省高校自然科学基金(14KJB310007);南京医科大学科技发展基金(2013NJMU015);江苏高校优势学科建设工程资助项目(13JC009);江苏省大学生创新训练计划项目(201410312042Y)


miR-372 regulates angiogenic gene AGGF1 expression and inhibits angiogenesis
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    摘要:

    目的:研究miR-372(miR-372)对新血管生成因子AGGF1基因的表达调控-方法:通过生物信息学预测找到可能靶向调控AGGF1基因的候选miRNA;通过荧光素酶报告基因实验-荧光定量聚合酶链反应(qPCR)和Western blot检测等实验证实miR-372对AGGF1基因表达的靶向调控;通过体外血管内皮细胞成管实验检测miR-372对血管生成的影响-结果:生物信息学预测显示miR-372靶向AGGF1;荧光素酶报告基因实验证实miR-372可通过结合AGGF1基因3′-非翻译区(3′-untranslational region,3′-UTR)而抑制其表达;qPCR和Western blot实验分别表明miR-372在mRNA水平和蛋白水平下调AGGF1表达;体外血管生成实验表明miR-372所介导的AGGF1表达下调可明显抑制血管生成-结论:miR-372可通过靶向结合AGGF1基因的3′-UTR下调其表达,并抑制内皮细胞血管生成能力-

    Abstract:

    Objective:To explore the regulation of angiogenic gene AGGF1 expression by miR-372. Methods: Bioinformatic prediction was performed to find the candidate microRNAs that may target AGGF1. Luciferase assays using AGGF1 3′-untranslational region (3′-UTR) reporters were performed to verify the binding of miR-372 to the AGGF1 3′-UTR; Real-time quantitative PCR (qPCR) and Western Blot assays were performed to measure the endogenous AGGF1 expression in 293T cells and HUVEC cells at mRNA and protein levels affected by overexpression of miR-372. In vitro angiogenesis assay was performed to verify the effects of miR-372 mediated downregulation of AGGF1 on angiogenesis. Results: Bioinformatic assays indicated that miR-372 targeted AGGF1. Luciferase assays confirmed that miR-372 downregulated AGGF1 expression directly through the miR-372 binding sites on 3′-UTR of AGGF1. qPCR and Western Blot assays revealed overexpression of miR-372 by mimic reduced AGGF1 expression at both mRNA and protein levels. In vitro matrigel based angiogenesis assay showed that overexpression of miR-372 reduced angiogenesis by downregulation of AGGF1. Conclusion: miR-372 downregulates AGGF1 expression and reduces angiogenesis.

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戴 运,周必胜. miR-372调控血管生成因子AGGF1的表达并抑制血管生成[J].南京医科大学学报(自然科学版),2015,(7):933-937

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  • 收稿日期:2015-03-06
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  • 在线发布日期: 2015-07-12
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