ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2015, Vol. 46 ›› Issue (4): 534-541.doi: 10.11843/j.issn.0366-6964.2015.04.004

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Identification of APC as the Potential Target of miR-135a-5p in Pigs

CHEN Chen1,2,DENG Yuan1,REN Hui-bo1,ZHU Ji1,HU Xiong-gui1,JIANG Si-wen2*,PENG Ying-lin1*   

  1. (1.Hunan Institute of Animal and Veterinary Science,Changsha 410131,China;2.College of Animal Science and Technology,Huazhong Agricultural University,Wuhan 430070,China)
  • Received:2014-07-08 Online:2015-04-23 Published:2015-04-23

Abstract:

This research aimed to investigate the potential target genes related to adipose tissue development.The bioinformatics softwares were used to predict the target gene of ssc-miR-135a-5p,and it was found that the 3′ UTR of adenomatous polyposis coli (APC) gene had 2 putative binding sites with miR-135a-5p.The wild-type (APC 3′ UTR) and 3 mutated (APC 3′ UTR m1,APC 3′ UTR m2 and APC 3′ UTR m3) dual-luciferase vectors were constructed and then co-transfected into PK-15 cells with miR-135a-5p mimics or negative control (NC),respectively,and then the luciferase activity was detected.The results showed that miR-135a-5p mimics significantly decreased the luciferase activities of APC 3′ UTR and APC 3′ UTR m2 vectors (P<0.05),while APC 3′ UTR m1 and APC 3′ UTR m3 completely abolished the inhibitory effects of miR-135a-5p,which demonstrated that miR-135a-5p mainly binded with the first site in 3′ UTR of APC gene.Further analysis revealed that miR-135a-5p inhibited the levels of endogenous APC mRNA and protein in porcine preadipocytes,which indicate that the target gene of ssc-miR-135a-5p is APC gene and regulates its expression.Taken together,this study will provide new insight into the biological function of ssc-miR-135a-5p and its mechanism in regulating adipogenesis.

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