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Keiichi Aomatsu, Tokuzo Arao, Kosuke Abe, Koji Sugioka, Aya Kodama, Hiroshi Mishima, Kazuto Nishio, Yoshikazu Shimomura; Slug Is Upregulated During Wound Healing And Downregulates Tp63 In Corneal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2011;52(14):1979. doi: https://doi.org/.
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Involvement of epithelial mesenchymal transition (EMT) in corneal wound healing remains largely unclear. The purpose of study is to gain the insight into EMT and corneal wound healing.
Slug expression in murine corneal tissues was evaluated using fluorescence staining. Snail and Slug were retrovirally and stably introduced into SV40-immortalized human corneal epithelial cells (HCECs). All cells were cultured in DMEM/F12 medium supplemented with 15% fetal bovine serum and gentamicin at 37oC with 5% CO2. Real-time PCR was carried out using Thermal Cycler Dice ® RealTime System. GAPDH was used as an internal control to normalize and compare each sample. In western blot analysis, antibodies used in this study were as follows: anti-snail, slug, E-cadherin, vimentin, beta-actin, N-cadherin. Rhodamine-conjugated Fhalloidin, E-cadherin and vimentin antibody and 4’,6-diamidino-2-phenylindole (DAPI) were used for fluorescent cellular staining.
Slug was overexpressed in corneal epithelial cells during wound healing in vivo, while Snail was not upregulated. Therefore, we hypothesized that Slug plays key roles during wound healing. Overexpression of Snail or Slug induced the cellular morphology and cadherin switch in HCECs, demonstrating that these transcription factors were able to mediate clear EMT in HCECs. Cellular proliferation was suppressed in these transfectant. Regarding a stemness of corneal epithelial cells, overexpression of Snail or Slug mediated the downregulation of TP63. The result suggests that Snail or Slug expression may be involve in the stemness of corneal epithelial cells during wound healing.
We found that slug is upregulated during corneal wound healing and the expression lead to downregulate the TP63 in corneal epithelial cells. Our findings provide novel insight into EMT, corneal wound healing and the stemness of corneal epithelial cells.
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