miR-194-5p靶向调控BCKDHA基因的表达

doi:10.11843/j.issn.0366-6964.2017.08.005

摘要/Abstract

摘要：

旨在预测并验证调控支链氨基酸分解代谢的限速酶支链α-酮酸脱氢酶复合体（Branched-chain α-ketoacid dehydrogenase complex，BCKDH）中α亚基即BCKDHA基因表达的关键miRNA。本研究利用3个在线软件预测与该基因具有靶标关系的miRNAs。利用荧光定量PCR检测小鼠3T3-L1前脂肪细胞分化过程中和超表达后关键miRNA与BCKDHA的表达量。结果表明，miR-194-5p是调控BCKDHA表达的一个关键miRNA，其种子序列与BCKDHA 3'UTR第190~196位碱基完全互补。MiR-194-5p的表达随分化时间呈下降趋势，而BCKDHA mRNA则呈上升趋势。双荧光素酶报告结果也显示二者具有靶标关系，miR-194-5p下调BCKDHA的表达。超表达miR-194-5p后，BCKDHA的表达显著下调（P<0.05）。由此证明，miR-194-5p通过与BCKDHA基因3'UTR结合抑制其表达。

Abstract:

This study aimed to predict and validate the crucial miRNA regulating BCKDHA gene expression, where BCKDHA was α subunit of BCKDH (Branched-chain α-ketoacid dehydrogenase complex), which was a key rate-limiting enzyme in the catabolism of branched-chain amino acids. Three online softwares were used to predict miRNAs targeting BCKDHA. QPCR was used to detect expressions of the crucial miRNA and BCKDHA mRNA in 3T3-L1 cells during its differentiation and after over-expressing the crucial miRNA, respectively. The results showed that miR-194-5p was a key miRNA regulating BCKDHA expression with fully complementation between its seed sequence and the 190-196th bases of BCKDHA 3'UTR. The expression of miR-194-5p decreased while that of BCKDHA mRNA increased with differentiation. The target relationship was also validated by dual luciferase reporter results which showed that miR-194-5p negatively regulated the expression of BCKDHA gene. The expression of BCKDHA mRNA was significantly reduced(P<0.05) after over-expressing miR-194-5p. It is concluded that miR-194-5p inhibits the expression of BCKDHA by targeting its 3' UTR.

中图分类号:

S813.1

张莹姣, 乔利英, 景炅婕, 潘洋洋, 马元, 郭云利, 魏琼, 刘文忠. miR-194-5p靶向调控BCKDHA基因的表达[J]. 畜牧兽医学报, 2017, 48(8): 1416-1423.

ZHANG Ying-jiao, QIAO Li-ying, JING Jiong-jie, PAN Yang-yang, MA Yuan, GUO Yun-li, WEI Qiong, LIU Wen-zhong. miR-194-5p Targets BCKDHA Gene to Regulate Its Expression[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(8): 1416-1423.

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https://www.xmsyxb.com/CN/Y2017/V48/I8/1416

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