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[目的]在分析纳米二氧化钛引起巨噬细胞微小RNA(mi R)表达谱改变的基础上,选择与免疫通路相关的mi R-27b,探讨mi R-27b对巨噬细胞RAW264.7的生物学功能及其对靶基因磷脂酰肌醇-3-激酶调节亚基3(Pik3r3)的调控作用。[方法]用Diana-mir Path在线软件对mi R-27b进行生物学信息分析,并通过Targetscan软件预测靶基因位点。通过转染使RAW264.7细胞的mi R-27b表达上调,采用MTT、流式细胞技术观察转染后细胞增殖、细胞周期和细胞凋亡的变化,应用Western blot检测靶基因的蛋白表达水平。[结果]Diana-mir Path在线软件进行mi R-27b的信号通路及靶基因分析结果显示,mi R-27b参与的信号通路-ln P≥2.99(P<0.05)的有23个,其中得分最高的通路为T细胞受体信号通路(T cell receptor signaling pathway),得分为16.89。靶基因预测结果显示Pik3r3可能是其靶基因之一。转染mi R-27b模拟物后,转染组与阴性对照组相比较,光密度值无明显差异(P=0.725),细胞周期也无明显差异(G1、S和G2期分别为P=0.490,P=0.088,P=0.323),但细胞凋亡率明显增高(P=0.023)。此外,转染组Pik3r3蛋白表达量无明显差异(P>0.05)。[结论]mi R-27b可促进RAW264.7细胞凋亡,其调控机制尚需进一步研究。
[Objective] To analyze the mi R-27b related to immune pathway based on the analysis of the change of micro-RNA (mi R) expression profile of macrophages by nano-TiO 2 and to investigate the biological effects of mi R-27b on macrophage RAW264.7 Function and its regulation of the target gene phosphatidylinositol 3-kinase regulatory subunit 3 (Pik3r3). [Methods] The biological information of mi R-27b was analyzed by online software Diana-mir Path, and the target gene locus was predicted by Targetscan software. The expression of mi R-27b in RAW264.7 cells was up-regulated by transfection. The proliferation, cell cycle and apoptosis were observed by MTT and flow cytometry. The protein expression of target gene was detected by Western blot. [Results] The results of mi R-27b signaling pathway and target gene analysis by online software Diana-mir Path showed that mi R-27b involved 23 signaling pathways with -ln P≥2.99 (P <0.05) Of the pathways for the T cell receptor signaling pathway (T cell receptor signaling pathway), a score of 16.89. Prediction of target genes showed that Pik3r3 may be one of its target genes. After transfection with mi R-27b mimics, there was no significant difference in optical density between transfection group and negative control group (P = 0.725) and no significant difference in cell cycle (P = 0.490 for G1, S and G2 phases , P = 0.088, P = 0.323), but the apoptosis rate was significantly higher (P = 0.023). In addition, there was no significant difference in Pik3r3 protein expression (P> 0.05). [Conclusion] mi R-27b can promote the apoptosis of RAW264.7 cells, and its regulatory mechanism needs further study.