Li Yanyan, Wang Xingguo, Yu Jianfeng, Shao Fang, Zhang Yanping, Lu Xiangyun, Gu Zhiliang
Department of Life Science and Technology, Changshu Institute of Technology, Changshu, Jiangsu, 215500, China.
Department of Life Science and Technology, Changshu Institute of Technology, Changshu, Jiangsu, 215500, China Jiangsu Institute of Poultry Science, Yangzhou, Jiangsu 225125, China.
Poult Sci. 2016 May;95(5):1145-50. doi: 10.3382/ps/pew039. Epub 2016 Mar 4.
As the most abundant microRNA (miRNA) in the liver, miR-122 plays important roles in the growth and development of liver, lipid metabolism, and liver diseases. Vanin 1 (VNN1) plays an important role in hepatic lipid metabolism, and VNN1 may serve as a potential therapeutic target for the treatment of metabolic diseases caused by overactivated gluconeogenesis. In our previous RNA-seq study, we found the expression of VNN1 increased significantly when the expression of miR-122 (gga-miR-122-5p) was knocked down in primary chicken hepatocytes. In this study, we verified this result by real-time qRT-PCR, and we also found that the chicken VNN1 was highly expressed in the liver. By bioinformatics analyses, we found the 3'UTR of VNN1 contained sequences completely complementary to the nucleotides 1 to 8 of miR-122. Co-transfection and dual-luciferase reporter assays showed that overexpression of miR-122 decreased the expression of luciferase reporter gene linked to the 3'UTR of chicken VNN1 in the Chinese hamster ovary cells (P<0.01), and the decrease was further demonstrated to be dependent on the predicted miR-122 binding sites by site mutation analyses. These results further support miR-122 as a negative regulator of VNN1 expression in chicken hepatocytes. Overall, this study suggests that miR-122 might play an important role in lipid metabolism in the chicken liver by negatively regulating the expression of the VNN1 gene.
作为肝脏中含量最丰富的微小RNA(miRNA),miR-122在肝脏的生长发育、脂质代谢及肝脏疾病中发挥着重要作用。泛酰巯基乙胺酶1(VNN1)在肝脏脂质代谢中起重要作用,且VNN1可能作为治疗因糖异生过度激活引起的代谢性疾病的潜在治疗靶点。在我们之前的RNA测序研究中,我们发现,在原代鸡肝细胞中敲低miR-122(gga-miR-122-5p)的表达时,VNN1的表达显著增加。在本研究中,我们通过实时定量逆转录聚合酶链反应(qRT-PCR)验证了这一结果,并且我们还发现鸡VNN1在肝脏中高表达。通过生物信息学分析,我们发现VNN1的3'非翻译区(UTR)包含与miR-122的第1至8个核苷酸完全互补的序列。共转染和双荧光素酶报告基因检测表明,在中华仓鼠卵巢细胞中,miR-122的过表达降低了与鸡VNN1的3'UTR相连的荧光素酶报告基因(P<0.01),并且通过位点突变分析进一步证明这种降低依赖于预测的miR-122结合位点。这些结果进一步支持miR-122作为鸡肝细胞中VNN1表达的负调节因子。总体而言,本研究表明miR-122可能通过负调节VNN1基因的表达在鸡肝脏脂质代谢中发挥重要作用。