Research Paper Volume 11, Issue 20 pp 8969—8981

Expression and significance of miR - 20b in retinal photoreceptor cells exposed to PCB1254

Xin Zhang 1, 2, 3, *, , Qingyu Zhang 3, 4, *, , Yue Jiang 1, 2, 3, *, , Shuchun Zhang 1, 2, 3, , Qin Hong 1, 2, 3, , Xirong Guo 1, 2, 3, , Xia Chi 1, 2, 3, , Meiling Tong 1, 2, 3, ,

  • 1 Department of Child Health Care, The Affiliated Obstetrics and Gynecology Hospital of Nanjing Medical University, Nanjing 210004, China
  • 2 Department of Child Health Care, Nanjing Maternity and Child Health Care Hospital, Nanjing 210004, China
  • 3 Department of Pediatrics, Nanjing Medical University, Nanjing 210029, China
  • 4 Department of Child Health Care, Northwest Women’s and Children’s Hospital, Xian 710061, China
* Equal contribution

received: March 27, 2019 ; accepted: October 5, 2019 ; published: October 16, 2019 ;

https://doi.org/10.18632/aging.102360
How to Cite

Copyright © 2019 Zhang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Previous studies have shown that PCB1254 has an adverse effect on zebrafish retinal development, but the basic mechanism behind it is not clear. The purpose of this study was to investigate the molecular mechanisms of PCB-induced retinal dysplasia. RT-qPCR, immunoblotting, HE staining and immunofluorescence were adopted to detect the expression at mRNA and protein level. Functional experiments were carried out in 661w cells including CCK-8 assay, caspase-3 assay, and the flow cytometry, while the functional role of miR - 20b was further investigated by using the zebrafish model. The result showed that PCB1254 exposure inhibited cell proliferation and increased the apoptosis of the 661w cells, and the dose–response relationship between the retinal development-related genes (SWS1, CRX, Rho), miR-20b expression and PCB1254 exposure was also discovered. We confirmed that miR-20b targeted FGF2 and GRB2 by constructing a dual luciferase reporter gene and suppressed the cell function as well as PCB1254. In the miR-20b overexpression zebrafish model, we found abnormal retinal morphology characterized by sparse and irregular photoreceptor cells and the thick photoreceptor cell layers. Our results demonstrate for the first time that PCBs target the MAPK/ERK signaling through miR-20b, affecting retinal cell development and leading to visual impairment.

Abbreviations

PCBs: Polychlorinated biphenyls; miRNA: microRNA; OMRs: optomotor responses; ERKs: Extracellular signal-regulated kinases; hpf: hours post fertilization; 661w cells: photoreceptor cell lines; RT-qPCR: Real-time quantitative polymerase chain reaction.