雷帕霉素和3-甲基腺嘌呤对RAW264.7细胞中8种miRNAs表达的影响

发布时间：2018-05-22 07:33

本文选题：自噬 + miRNAs　；参考：《免疫学杂志》2017年09期

【摘要】：目的探究雷帕霉素(Rapamycin,Rapa)和3-甲基腺嘌呤(3-methyladenine,3-MA)处理对RAW264.7细胞中miR-181c,miR-101b和miR-1192等8种自噬相关miRNAs的表达水平,为研究miRNAs在细胞自噬中的功能提供理论基础。方法生物信息学预测分析与细胞自噬相关的miRNAs并构建miRNAs调控细胞自噬的网络图;分别用Rapa和3-MA对RAW264.7细胞做不同条件处理,通过实时荧光定量PCR(quantitative real-time PCR,q RT-PCR)检测不同条件处理后RAW264.7细胞中miR-181b、miR-181c、miR-101b、miR-1192、miR-362-3p、miR-590-3p、miR-875-5p和miR-335-5p的相对表达量。结果生物信息学预测结果显示8种miRNAs可能分别靶向作用于细胞自噬通路中多个关键蛋白,对细胞自噬起到促进或抑制作用;与正常对照组相比,miR-181c、miR-101b、miR-1192、miR-362-3p、miR-335-5p、miR-875-5p在Rapa处理RAW264.7细胞组中表达显著下调(P0.05),而在3-MA处理组则都显著上调(P0.05);miR-181b在Rapa组表达显著上调,在3-MA组表达下调(P0.05);miR-590-3p在Rapa组和3-MA组都无显著性变化(P0.05)。结论 miR-181c、miR-101b等7种miRNAs可能在RAW264.7巨噬细胞自噬过程中发挥重要调控作用。
[Abstract]:Objective to investigate the expression of miR-181 cmmiR-101b and miR-1192 in RAW264.7 cells treated with rapamycin (Rapa) and 3-methyladenine 3-MA), and to provide a theoretical basis for the study of the function of miRNAs in autophagy. Methods Bioinformatics was used to predict and analyze the miRNAs associated with autophagy and to construct a network map of miRNAs regulating autophagy. Rapa and 3-MA were used to treat RAW264.7 cells under different conditions, respectively. The relative expression levels of miR-181bsimiR-181cnmiR-101btmiR-1192pmmiR-362-3pmmiR-590-3pmmiR-875-5p and miR-335-5p in RAW264.7 cells were detected by real-time fluorescence quantitative PCR(quantitative real-time PCRQ RT-PCR. Results the bioinformatics prediction results showed that the eight kinds of miRNAs might target several key proteins in the autophagy pathway and promote or inhibit the autophagy. Compared with normal control group, miR-181 cmiR-101b miR-1192 miR-362-3 pnmiR-335-5 pnmiR-875-5p significantly down-regulated P0.05P in RAW264.7 cells treated with Rapa, while in 3-MA group, the expression of P0.05miR-181b was significantly up-regulated in Rapa group, but no significant change was found in 3-MA group. Conclusion miR-181 cu miR-101b and other seven miRNAs may play an important role in the process of RAW264.7 macrophage autophagy.
【作者单位】：宁夏医科大学临床医学院医学检验系;宁夏医科大学基础医学院医学遗传系与细胞生物学系;
【基金】：国家自然科学基金(31472168) 宁夏科技创新领军人才项目(2015年)
【分类号】：R392

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