College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu225002, China.
Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou225009, Jiangsu Province, China.
J Dairy Res. 2020 May;87(2):232-238. doi: 10.1017/S0022029920000229. Epub 2020 Apr 16.
In this research paper we filter and verify miRNAs which may target silent information regulator homolog 2 (SIRT2) gene and then describe the mechanism whereby miRNA-212 might regulate lipogenic genes in mammary epithelial cell lines via targeting SIRT2. Bioinformatics analysis revealed that the bovine SIRT2 gene is regulated by three miRNAs: miR-212, miR-375 and miR-655. The three miRNAs were verified and screened by qRT-PCR, western blot, and luciferase multiplex verification techniques and only miR-212 was shown to have a targeting relationship with SIRT2. The results of co-transfecting miR-212 and silencing RNA (siRNA) showed that by targeting SIRT2, miR-212 can regulate the expression of fatty acid synthetase (FASN) and sterol regulatory element binding factor 1 (SREBP1) but not peroxisome proliferator-activated receptor gamma (PPARγ). Measurement of triglyceride (TAG) content showed that miR-212 increased the fat content of mammary epithelial cell lines. The study indicates that miR-212 could target and inhibit the expression of the SIRT2 gene to promote lipogenesis in mammary epithelial cell lines.
在本研究论文中，我们筛选并验证了可能靶向沉默信息调节因子同源物 2（SIRT2）基因的 miRNAs，然后描述了 miRNA-212 可能通过靶向 SIRT2 调节乳腺上皮细胞系中脂肪生成基因的机制。生物信息学分析表明，牛 SIRT2 基因受三种 miRNAs 的调控：miR-212、miR-375 和 miR-655。通过 qRT-PCR、western blot 和荧光素酶多重验证技术验证和筛选了这三种 miRNAs，只有 miR-212 与 SIRT2 具有靶向关系。共转染 miR-212 和沉默 RNA（siRNA）的结果表明，通过靶向 SIRT2，miR-212 可以调节脂肪酸合成酶（FASN）和固醇调节元件结合蛋白 1（SREBP1）的表达，但不能调节过氧化物酶体增殖物激活受体 γ（PPARγ）的表达。甘油三酯（TAG）含量的测量表明，miR-212 增加了乳腺上皮细胞系的脂肪含量。该研究表明，miR-212 可以靶向并抑制 SIRT2 基因的表达，从而促进乳腺上皮细胞系中的脂肪生成。
