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利用生物信息学方法预测出miR-224可能的靶基因为ACADM、ACAT1、ALDH2和LPL,然后用实时荧光定量法检测了miR-224与其靶基因在奶牛乳腺上皮细胞中的表达量。结果显示:(1)miR-224的相对表达量。转染shNc组表达量低于转染mimics组而高于inhibitor组。(2)靶基因荧光定量结果表明,ACADM、ALDH2和LPL基因转染shNc组表达量低于转染inhibitor组而高于mimics组。(3)经SPSS22.0相关性分析表明:在乳腺上皮细胞中,ACADM、ALDH2和LPL基因的表达量与miR-224的表达量呈显著负相关,ACAT1基因与miR-224没有显著联系。初步验证说明,ACADM、ALDH2和LPL基因可能是miR-224的靶基因,为进一步探索miR-224在奶牛乳牛乳腺上皮细胞发挥的作用提供了一定的理论依据和研究基础。
Bioinformatics methods were used to predict the possible target genes of miR-224 as ACADM, ACAT1, ALDH2 and LPL. The expression of miR-224 and its target gene in mammary epithelial cells was detected by real-time fluorescence quantitative assay. The results showed: (1) The relative expression of miR-224. Transfection of shNc group was lower than the transfected mimics group and higher than the inhibitor group. (2) Fluorescence quantification of target genes showed that expression of ACADM, ALDH2 and LPL genes in shNc group was lower than that in transfected inhibitor group and mimics group. (3) SPSS22.0 correlation analysis showed that: in the mammary epithelial cells, ACADM, ALDH2 and LPL gene expression was negatively correlated with the expression of miR-224, ACAT1 gene and miR-224 was not significantly linked. Preliminary validation indicated that ACADM, ALDH2 and LPL genes may be the target genes of miR-224, which provide some theoretical basis and research basis for further exploring the role of miR-224 in dairy cow mammary epithelial cells.