Effects of rapamycin and 3-methyladenine on the expression of eight miRNAs in RAW264.7 cells

XU Xiangrong; MA Zhanbing; JIANG Dan; QU Yuliang; WANG Xian; FANG Yuan; GUO Le; XU Guangxian

doi:10.13431/j.cnki.immunol.j.20170141

Abstract：This study was performed to determine the expression levels of miR-181c,miR-101b and miR-1192 etc.in mouse RAW264.7 cells and provide a theoretical basis for exploring the function of miRNAs in macrophage autophagy.Bioinformatics analysis was used to predict autophagy-related miRNAs and establish miRNAs-regulated network of autophagy.The relative expression of miR-181b,miR-181c,miR-101b,miR-1192,miR-362-3p,miR-590-3p,miR-875-5p and miR-335-5p were detected by qRT-PCR in RAW264.7 cell models pre-treated with different conditions of Rapamycin (Rapa) or 3-methyladenine (3-MA).The results of bioinformatics showed that eight miRNAs could target key proteins in autophagy pathway,which may facilitate or inhibit autophagy function.Compared with the normal control,the expression of miR-181c,miR-101b,miR-1192,miR-362-3p,miR-335-5p and miR-875-5p in RAW264.7 cells was significantly down-regulated after Rapa treatment (P＜0.05),while markedly up-regulated in the 3-MA treatment group (P＜0.05);however,the opposite expression level variation of miR-181b was observed under the same conditions as above (P＜0.05).The expression of miR-590-3p had no change after any treatments (P＞0.05).In conclusion,our research suggest that,except for miR-590-3p,other seven miRNAs present remarkably differential expression and thus play critical regulatory roles in RAW264.7 mouse macrophage autophagy.