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A miR-511-binding site SNP in the 3′UTR of IGF-1 gene is associated with proliferation and apoptosis of PK-15 cells

  • Wenzhen Wei
  • Gang Wang
  • Yunyun Cheng
  • Rui Yang
  • Jie Song
  • Shan Huang
  • Haoyang Li
  • Hongwei Geng
  • Hao Yu
  • Songcai LiuEmail author
  • Linlin HaoEmail author
Article
  • 27 Downloads

Abstract

Insulin-like growth factor-1 (IGF-1) is a functional candidate gene for pig growth and development due to its crucial role in the growth axis of growth hormone-IGF-1. Considering that the 3′ untranslated region (3′UTR) of gene may affect its expression, we analyzed the effect of a single-nucleotide polymorphism (SNP) (rs34142920, c.674C > T) on gene expression, cell proliferation, and apoptosis and the possible related molecular mechanisms in PK-15 cells. The SNP was found in the 3′UTR of IGF-1 in Bama Xiang pig in previous investigations. Results showed that the SNP was located at the target site binding to microRNA (miR-511). The 3′UTR of IGF-1 gene with C allele significantly downregulated the expression of IGF-1 gene compared with that of the gene with T allele by luciferase assay. miR-511 was transfected into porcine kidney cell line (PK-15 cells) to reveal its effects on cells and whether or not it targets IGF-1. The expression levels of IGF-1 at mRNA and protein levels were remarkably downregulated. miR-511 significantly inhibited cell proliferation and promoted cell apoptosis by downregulating the phosphorylation level of AKT and ERK1/2. This finding confirmed that miR-511 inhibits proliferation and promotes apoptosis by downregulating the IGF-1 in PK-15 cells.

Keywords

IGF-1 SNP miR-511 Proliferation Apoptosis 

Notes

Funding information

This work was supported by the National Natural Science Foundation of China (31672514 and 31772699) and the Jilin Scientific and Technological Development Program (20170101024JC).

Compliance with ethical standards

All of the protocols in this study were approved by the Ethics Committee on the Use and Care of Animals at Jilin University (Changchun, China) and were in compliance with the National Institute of Health Guide for the Care and Use of Laboratory Animals.

Conflict of interest

The authors declared that they have no conflicts of interest.

Supplementary material

11626_2019_329_Fig5_ESM.png (369 kb)
Fig. S1

Healthy PK-15 cell morphology. Images obtained using a 10x eyepiece with 4x, 10x, 20x and 40x objective lenses (Figs. A, B, C and D, respectively) were taken 1 day after inoculation (PNG 368 kb)

11626_2019_329_MOESM1_ESM.tif (791 kb)
High Resolution Image (TIF 791 kb)
11626_2019_329_Fig6_ESM.png (801 kb)
Fig. S2

PCR product sequencing and NCBI database alignment analysis. (A) Sanger sequencing analysis of the 3′UTR product of IGF-1 amplified in PK-15 cells. (B) NCBI database alignment analysis of the results in (A) (PNG 800 kb)

11626_2019_329_MOESM2_ESM.tif (6 mb)
High Resolution Image (TIF 6093 kb)
11626_2019_329_Fig7_ESM.png (858 kb)
Fig. S3

Description of PK-15 cells (PNG 857 kb)

11626_2019_329_MOESM3_ESM.tif (6.7 mb)
High Resolution Image (TIF 6825 kb)

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Copyright information

© The Society for In Vitro Biology 2019

Authors and Affiliations

  • Wenzhen Wei
    • 1
  • Gang Wang
    • 1
  • Yunyun Cheng
    • 1
    • 2
  • Rui Yang
    • 1
  • Jie Song
    • 1
  • Shan Huang
    • 1
  • Haoyang Li
    • 1
  • Hongwei Geng
    • 1
  • Hao Yu
    • 1
  • Songcai Liu
    • 1
    • 3
    Email author
  • Linlin Hao
    • 1
    Email author
  1. 1.College of Animal ScienceJilin UniversityChangchunChina
  2. 2.College of Animal ScienceSouth China Agricultural UniversityGuangzhouChina
  3. 3.Five-Star Animal Health Pharmaceutical Factory of Jilin ProvinceChangchunChina

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