Role of miR‑23a/Zeb1 negative feedback loop in regulating epithelial‑mesenchymal transition and tumorigenicity of intraocular tumors

Wang,Yanyan; Luo,Yunna; Guan,Wenying; Zhao,Haixia

doi:10.3892/ol.2018.8940

Role of miR‑23a/Zeb1 negative feedback loop in regulating epithelial‑mesenchymal transition and tumorigenicity of intraocular tumors

Authors:
- Yanyan Wang
- Yunna Luo
- Wenying Guan
- Haixia Zhao
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Affiliations: Department of Ophthalmology, Affiliated Hospital of Inner Mongolia Medical University, Hohhot, Inner Mongolia 010059, P.R. China
Published online on: June 11, 2018 https://doi.org/10.3892/ol.2018.8940
Pages: 2462-2470

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Abstract

Role of the two‑way negative feedback regulation channel formed by miR‑23a and Zeb1 in epithelial‑mesenchymal transition (EMT), tumorigenic ability, and migration and metastasis capacity of the intraocular malignant tumor cells was investigated. Molecular biological methods such as real time-quantitative PCR (RT-qPCR), immunoblotting method, and immunofluorescence were used to detect the expression levels of mRNA and protein in the Zeb1 factor in OCM‑1, WERI‑RB1, and Y79 cells before and after miR‑23a transfection. Transwell cells were used to detect the in vitro membrane permeation and migration ability in OCM‑1, WERI‑RB1, and Y79 cells (non‑transfection group, blank control transfection group, mimic transfection group, inhibitor transfection group). The results revealed that the relative expression of miR‑23a in the cells in the miR‑23a mimic transfection group increased significantly compared with that in the control group (p<0.05). There were significant differences in the relative expression of mRNA between the mimic transfection and control group (p<0.05). RT‑qPCR detection showed that the relative expression of mRNA of the epithelial‑labeled factor E‑cadherin increased significantly in the miR‑23a mimics group (p<0.05). Expression of the protein E‑cadherin increased while the expression of the mesenchyme‑labeled proteins of vimentin and N‑cadherin decreased in the mimics group. Zeb1 has a negative feedback effect on miR‑23a. They can form a negative feedback loop. The results showed that miR‑23a and Zeb1 form a bidirectional inhibitory negative feedback loop, which plays an important role in regulating EMT. In conclusion, the significant changes in the mesenchymal phenotype of the stable strains with Zeb1 overexpressed in the OCM‑1 cells cannot be completely explained with the changes in cytoskeleton caused by EMT.

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