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目的:研究miR-142-3p对自噬相关基因ATG4c的靶向调控作用,探究miR-142-3p影响RAW264.7细胞自噬途径的作用机制。方法:生物信息学软件分析miR-142-3p的靶基因为ATG4c,构建pMIR-Report-ATG4c和pMIR-Report -ATG4c mut重组质粒,双荧光素酶报告系统、qRT-PCR、Western blot 验证 miR-142-3p 与 ATG4c 的靶向作用;将做不同处理的RAW264.7细胞分为4组:正常细胞作为对照、50 ng/ml雷帕霉素作用2 h、EBSS饥饿作用12 h、10 nmol/L的3-甲基腺嘌呤(3-MA)作用12 h后,实时荧光定量PCR(qRT-PCR)检测miR-142-3p不同干预组中的相对表达情况;将miR-142-3p mimics、miR-142-3p inhibitor及miR-142-3 p control分别转染到RAW264.7细胞中,检测miR-142-3p和LC3Ⅱ的相对表达。结果:双荧光素酶报告系统、qRT-PCR、Western blot验证miR-142-3p通过靶向作用于ATG4c的3′-UTR抑制其表达;与对照组相比,雷帕霉素和饥饿处理的RAW264.7细胞miR-142-3p明显上调,而3-MA处理组miR-142-3p明显下调;与miR-142-3p control组相比,转染miR-142-3p mimics组中LC3Ⅱ蛋白表达显著下调,而miR-142-3p inhibitor 组中表达显著上调。结论:miR-142-3p通过靶向调控自噬相关基因ATG4c,参与RAW264.7小鼠巨噬细胞自噬的调控。“,”Objectiev:To study the mechanism of miR-142-3p regulated Autophagy Association Gene (ATG4c),to explore autophagy pathway in RAW264.7 macrophages.Methods: To predict the target genes of miR-142-3p by Bioinformatics Software—ATG4c.Then to establish pMIR-Report-ATG4c and pMIR-Report-ATG4 c mut recombinant plasmid,in order to confirm the regulatory relation between miR-142-3p and ATG4c through dual luciferase assay ,qRT-PCR,Western blot.Four groups was assigned by different treatment,they were normal cells control group ,2 h 50 ng/ml rapamycin treated group ,12 h EBSS hunger treated group and 12 h 10 nmol/L 3-methyl adenine (3-MA) treated group,to detect miR-142-3p relative expression in RAW264.7 cell line by quantitative Real-Time PCR ( qRT-PCR).MiR-142-3p mimics, miR-142-3p inhibitor and miR-142-3p control was transfected in RAW264.7 cells respectively,the relative expression of miR-142-3p and LC3Ⅱ expression was examined by qRT-PCR and Western blot.Res ults: It revealed the target gene (ATG4c) of miR-142-3p by Dual luciferase assay,qRT-PCR and Western blot;Compared with the control group,the expression of miR-142-3p was obviously up-reguated from the rapamycin and hunger treated groups in RAW 264.7cell line, on the contrary,miR-142-3p was obviously down regulated from the 3-MA treated group in RAW264.7cell line;meanwhile,compared with the control group , LC3Ⅱ expression was significantly lower from miR -1423-p mimics group;however , LC3Ⅱ expression was significantly higher from miR-142-3p inhibitor group.Conclusion: MiR-142-3p can target associated genes ATG4c to regulate autophagy in RAW264.7 cell line.