Chicken GHR antisense transcript regulates its sense transcript in hepatocytes

Gene. 2019 Jan 15:682:101-110. doi: 10.1016/j.gene.2018.10.001. Epub 2018 Oct 5.

Abstract

An increasing number of evidences indicated that long noncoding RNAs (LncRNAs) regulate a variety of biological progresses via different mechanisms. Our previous study had identified a chicken growth hormone receptor (GHR) antisense transcript (GHR-AS) which regulated GHR sense transcript (GHR-S) in LMH cells. In the present study, roles of GHR-AS and its regulatory mechanism were analyzed in chicken hepatocytes. The expression patterns of liver GHR-S, GHR-AS and Let-7b ascended with the development of chicken. The hepatocytes proliferation was promoted and more cells entered into DNA synthesis (S) phase when GHR-AS was overexpressed while the cell proliferation was slowed and fewer cells were in S phase when GHR-AS was interfered. Meanwhile, the GHR-S increased when we overexpressed GHR-AS while it reduced when GHR-AS was inhibited. The S1 Nuclease protection assay indicated that GHR-S and GHR-AS formed RNA duplex via GHR-S 3' untranslation regon (3'UTR). In hepatocytes or LMH cells, the half-time of GHR-S showed a delayed trend when GHR-AS or GHR-AS 5' untranslation regon (5'UTR) was overexpressed. Furthermore, the level of GHR-S can be decreased by Let-7b mimics whereas it was partially rescued when co-transfected pGHR-AS or pGHR-AS 5'UTR with Let-7b mimics. Based on our findings, GHR-AS affected hepatocytes proliferation and improved GHR-S stability possibly by forming RNA duplex between GHR-S and GHR-AS, competing with Let-7b.

Keywords: Double strand RNA; Growth hormone receptor; Natural antisense transcript; Stability.

MeSH terms

  • 3' Untranslated Regions
  • 5' Untranslated Regions
  • Animals
  • Cell Proliferation
  • Chick Embryo
  • Chickens / genetics
  • Gene Expression Profiling
  • Gene Expression Regulation*
  • Hepatocytes / cytology
  • Hepatocytes / metabolism*
  • Liver / metabolism
  • Oligonucleotides, Antisense / genetics*
  • Receptors, Somatotropin / genetics*
  • Transfection

Substances

  • 3' Untranslated Regions
  • 5' Untranslated Regions
  • Oligonucleotides, Antisense
  • Receptors, Somatotropin