Di-san junyi daxue xuebao (Feb 2020)

MiR-200b inhibits corneal angiogenesis in rats by regulating PIK3CA/AKT pathway

  • LI Xue,
  • CAO Lang,
  • GAN Min,
  • ZHOU Shanbi

DOI
https://doi.org/10.16016/j.1000-5404.201909141
Journal volume & issue
Vol. 42, no. 4
pp. 407 – 413

Abstract

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Objective To investigate the role of microRNA-200b (miR-200b) in corneal neovascularization (CNV) in rats and explore the possible mechanisms. Methods Twenty-four female SD rats were subjected to corneal suture to induce CNV in one eye, with the other eye as the normal control. On the 4th, 7th and 14th days after the suture, CNV was observed under a slit lamp, and the structure and inflammatory cell infiltration in the corneal tissue were observed using HE staining. On day 14 after the suture, the expression levels of miR-200b and vascular endothelial growth factor (VEGF) mRNA in the cornea were detected using qRT-PCR. Human umbilical vein endothelial cells (HUVECs) were transfected via liposome with a miR-200b mimic or a negative control sequence (miR-NC), and the cell proliferation, migration and tube formation abilities were evaluated using CCK-8 assay, Transwell chamber assay and Matrigel assay, respectively. The expression levels of VEGF and PIK3CA/AKT proteins in the cells were detected by Western blotting. Results CNV models were successfully established by corneal suture in the rats. HE staining showed that corneal suture resulted in disordered alignment of the corneal tissue and progressively increased inflammatory cell infiltration on days 4, 7 and 14 after the suture (P < 0.05). RT-PCR showed that miR-200b expression decreased and VEGF expression increased significantly in the corneal tissue on day 14 after the suture (P < 0.01). Compared with the cells without transfection or transfected with miR-NC, the HUVECs transfected with the miR-200b mimic exhibited significantly attenuated proliferation, migration and lumen formation abilities (P < 0.05) with also significantly lowered expression of VEGF, PIK3CA and p-AKT proteins (P < 0.05). Conclusion miRNA-200b can significantly inhibit the proliferation, migration and tube formation of HUVECs, and may participate in the occurrence and development of CNV by inhibiting the PIK3CA/AKT pathway.

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