论文部分内容阅读
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(P<0.05),而在3-MA处理组则都显著上调(P<0.05);miR-181b在Rapa组表达显著上调,在3-MA组表达下调(P<0.05);miR-590-3p在Rapa组和3-MA组都无显著性变化(P>0.05)。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。
OBJECTIVE: To investigate the effects of Rapamycin (Rapa) and 3-methyladenine (3-MA) on the expression of miR-181c, miR-101b and miR-1192 in RAW264.7 cells Related miRNAs expression levels provide a theoretical basis for studying the function of miRNAs in autophagy. Methods Bioinformatics predicts the miRNAs associated with autophagy and constructs a network map of miRNAs regulating autophagy. RAW264.7 cells were treated with different conditions by Rapa and 3-MA, respectively, and then quantitative real- The expression of miR-181b, miR-181c, miR-101b, miR-1192, miR-362-3p, miR-590-3p and miR-875- 5p and miR-335-5p relative expression levels. Results Bioinformatics prediction showed that 8 kinds of miRNAs could target multiple key proteins in autophagy pathway respectively, which could promote or inhibit autophagy. Compared with normal control group, miR-181c and miR- 101b, miR-1192, miR-362-3p, miR-335-5p and miR-875-5p were significantly down-regulated in Rapa-treated RAW264.7 cells (P <0.05) (P <0.05). MiR-181b was up-regulated in Rapa group and down-regulated in 3-MA group (P <0.05). MiR-590-3p showed no significant change in Rapa group and 3-MA group P> 0.05). Conclusion Seven miRNAs such as miR-181c and miR-101b may play important roles in the autophagy of RAW264.7 macrophages.