利用LPS刺激RAW264.7细胞,在不同的时间段检测到miR-17-5p和miR-20a表达水平的变化.利用基因克隆技术构建miR-17pcDNA3.1和miR-20a pcDNA3.1表达载体,利用电转的方法转染RAW264.7细胞,实时定量PCR方法检测RAW264.7细胞中miR-17和miR-20a的表达以确定转染率.LPS刺激4d后,进一步探讨miR-17和miR-20a对小鼠巨噬细胞表面抗原CD80和CD86表达的影响.结果显示,LPS刺激RAW264.7细胞检测到成熟miR-17-5p和miR-20a的表达先上升,至12h后表达开始下降,且克隆的miR-17pcDNA3.1和miR-20a pcDNA3.1质粒转染细胞后能产生成熟的miR-17-5p和miR-20a,LPS刺激CD80和CD86的上调能被miR-17和miR-20a抑制,说明miR-17和miR-20a能够调控抗原提呈细胞表面共刺激分子的表达.
The expression levels of miR-17-5p and miR-20a were measured in RAW264.7 in the presence of LPS at different time points.The results show that the expression of mature miR-17-5p and miR-20a is elevated and then dropped following LPS after 12 h.Meanwhile,it is also studied whether miR-17 and miR-20a could affect CD80 and CD86 expression in the presence of LPS.Upregulation of CD80 and CD86 is induced by LPS could be reversed by miR-17 and miR-20a transfection.It suggested that tumor associated miR-17 and miR-20a may play an important role in LPS-mediateing the expression of costimulatory molecules.