MicroRNA let-7g inhibited hypoxia-induced proliferation of PASMCs via G0/G1 cell cycle arrest by targeting c-myc

Life Sci. 2017 Feb 1:170:9-15. doi: 10.1016/j.lfs.2016.11.020. Epub 2016 Nov 24.

Abstract

Aims: Pulmonary hypertension (PH) is a proliferative disorder characterized by enhanced proliferation and suppressed apoptosis of intrapulmonary vascular smooth muscle cells. Recently, network-based bioinformatics have identified let-7 family, a tumor suppressive microRNA, regulate multiple interacting targets relevant to PH. However, the role of let-7 in vascular homeostasis in PH remains unknown. Thus, we wanted to investigate the role of let-7 in hypoxia-induced PASMCs proliferation and the underlying mechanism in hypoxic pulmonary hypertension (HPH).

Main methods: The male Sprague-Dawley (SD) rats were exposed to hypoxia (10% O2) for 21days to induce HPH. The expression of let-7 was determined by quantitative real-time polymerase chain reaction (qRT-PCR) and in situ hybridization. Primary rat PASMCs were exposed to hypoxia (3% O2). MTS and EDU were performed to evaluate PASMCs proliferation. The mRNA and protein expression of c-myc, Bmi-1 and p16 were determined by qRT-PCR and Western blotting, respectively. The functions of let-7g on PASMCs proliferation, c-myc, Bmi-1 and p16 expression were assessed by let-7g mimic and inhibitor transfection.

Key findings: Among let-7 family members, only let-7b and let-7g were significantly down-regulated in remodeled pulmonary artery in HPH rats. Furthermore, only let-7g level was decreased in hypoxic PASMCs. Either hypoxia or let-7g inhibitor stimulated proliferation of PASMCs, let-7g mimic inhibited hypoxia-induced PASMCs proliferation. C-myc was the target of let-7g in PASMCs. Transfect of let-7g mimic inhibited hypoxia-induced c-myc, Bmi-1 up-regulation and p16 down-regulation, which ultimately controls cell cycle progression.

Significance: Loss of inhibition on c-myc-Bmi-1-p16 signaling pathway by let-7g may lead to PASMCs proliferation and vascular remodeling in HPH.

Keywords: Cell cycle; Cell proliferation; HPH; Vascular remodeling; let-7; microRNA.

MeSH terms

  • Animals
  • Apoptosis
  • Cell Proliferation
  • Cyclin-Dependent Kinase Inhibitor p16 / metabolism
  • G1 Phase Cell Cycle Checkpoints
  • Homeostasis
  • Hypertension, Pulmonary / metabolism*
  • Hypertension, Pulmonary / pathology
  • Hypoxia*
  • In Situ Hybridization
  • Male
  • MicroRNAs / genetics*
  • Myocytes, Smooth Muscle / cytology
  • Polycomb Repressive Complex 1 / metabolism
  • Proto-Oncogene Proteins c-myc / metabolism*
  • Pulmonary Artery / metabolism*
  • Rats
  • Rats, Sprague-Dawley
  • Resting Phase, Cell Cycle
  • Signal Transduction
  • Vascular Remodeling / physiology

Substances

  • Bmi1 protein, rat
  • Cyclin-Dependent Kinase Inhibitor p16
  • MIRNLET7 microRNA, rat
  • MicroRNAs
  • Proto-Oncogene Proteins c-myc
  • Polycomb Repressive Complex 1