A SNP in the 3'UTR of the porcine IGF-1 gene interacts with miR-new14 to affect IGF-1 expression, proliferation and apoptosis of PK-15 cells

Abstract

The kidney of miniature pigs has been considered the most likely potential kidney source for patients needing kidney transplantation. Insulin-like growth factor 1 (IGF-1) is involved in regulating the growth of miniature pigs and inducing growth of kidneys. There are evidences showing that the SNPs in the 3'UTR of a gene may affect the gene expression by affecting the binding to a miRNA target site. In this study, one SNP (rs34142920) was screened in the IGF-1 3'UTR between 2 different body types of porcine breeds, Bama Xiang (BX) pigs, a miniature pig breed, and Large White (LW) pigs by sequencing. The secondary structure of the IGF-1 3'UTR mRNA containing the SNP in BX pigs is different from that of LW pigs. We then verified that there was a porcine miRNA (miR-new14) binding to this SNP in the 3'UTR of IGF-1 via cotransfecting the 3'UTR from the 2 breeds and miR-new14. We further found that the SNP downregulated mRNA and protein levels of IGF-1 by affecting the binding of miR-new14. To understand the function of miR-new14 in porcine kidney (PK-15) cells and its mechanism, cell proliferation and cell apoptosis assays were employed and results showed that proliferation viability of PK-15 cells was weakened and the apoptotic percentage of PK-15 cells was higher in the miR-new14 group. Porcine miRNA reduced the mRNA expression of AKT/ERK and protein levels of p-AKT/p-ERK. These results suggested that the expression of IGF-1 is influenced by this SNP and miR-new14 and that miR-new14 may suppress cell proliferation and promote cell apoptosis in PK-15 cells through regulating AKT and ERK signaling pathways, in which IGF-1 is involved.

Keywords: IGF-1; Porcine growth; Porcine kidney (PK-15) cells; SNP (rs34142920); miR-new14.