Abstract
Purpose :
Several improvements have been made to reduce the posterior capsule opacification (PCO) rate, however 10% of patients still undergo PCO. Selective covalent CDK7 inhibitor THZ1 has been proved to be a potential anti-tumor drug in various cancers, we hypothesize that THZ1 may reduce the rate of PCO via suppressing lens epithelial cells (LECs) epithelial-mesenchymal transition (EMT), as EMT is highly related to cancer initiation , metastasis as well as PCO.
Methods :
EMT was induced by TGFβ2 in Human SRA01/04 LECs, rabbit primary lens epithelial cells and whole rat lens culture semi-in vivo model, and then cultured with gradient concentration of THZ1. The expression level of mesenchymal markers including Snai1, ZEB1, Fibronectin, Collagen, N-cadherin, α-SMA as well as epithelial symbol markers E-cadherin and ZO-1 were detected by WB, RT-PCR and immunofluorescence analysis. Transparency of rat lens was photographed using a stereoscopic microcopy, scratch assay was also conducted to analysis the change of migratory ability in LECs after co-treatment of TGFβ2 and THZ1. In order to figure out the pathway THZ1 participated in suppressing EMT, we executed RNA sequencing and KEGG assay.
Results :
THZ1 suppressed the EMT progress induced by TGFβ2 in SRA01/04 LECs, rabbit primary lens epithelial cells and whole rat lens culture semi-in vivo model. The migration ability of LECs treated with TGFβ2 was stimulated, while depressed by THZ1 in a dose-dependent manner. THZ1 treatment also restored lens transparency reduced by TGFβ2 in rat lens. RNA-sequencing and KEGG analysis revealed that the THZ1 inhibits EMT through TGFβ/Smad and Notch signaling pathway, then we validated it by detecting the expression level of Notch pathway components and phosphorylation of Smad2/3.
Conclusions :
Our results are consistent with our hypothesis that as a potential antineoplastic compound, THZ1 strongly attenuated TGFβ2 induced EMT in LECs. Notch and TGFβ2/Smad pathway might be the agency for THZ1 to regulate EMT. These results indicate that THZ1 may function in preventing and curing PCO.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.