Long Non-coding RNA SNHG11 Regulates The Wnt/β-catenin Signaling Pathway Through Rho/ROCK In Trabecular Meshwork Cells

Purpose:Trabecular meshwork（TM）cell dysfunction is the main cause of elevated intraocular pressure（IOP）and glaucoma.The long non-codingRNA（lncRNA）small nucleolarRNA host gene 11（SNHG11）is associated with cell proliferation,apoptosis,and other cellular processes,but its biological role and mechanism in glaucoma remain unclear.In the present study,we investigated the role of SNHG11 in TM cells.Methods:1.SNHG11 expression in immortalized human TM（iHTM）cells,glaucomatous human TM（GTM₃）cells and a hypertension-dependent glaucoma mouse model was detected using quantitative real-time PCR analysis（q RT-PCR）.2.iHTM and GTM₃ cells were cultured and transfected with siRNA targeting SNHG11（si-SNHG11）,the transfection efficiency was detected by q RT-PCR.Transwell assays,q RT-PCR,western blotting,and CCK-8 assays were used to evaluate TM cell migration,apoptosis,autophagy,and survival.3.Wnt/β-catenin pathway activity was inferred from q RT-PCR,western blotting,immunofluorescence,and luciferase reporter and TOPFlash reporter assays in both two cells,glaucoma mouse model and transfected with siRNA in TM cells.4.Transfected siRNA and treated with Wnt pathway inhibitors or activators in iHTM cells,the Wnt/β-catenin signaling pathway activity was detected by q RT-PCR,western blotting,luciferase reporter and TOPFlash reporter assays.5.The expression of Rho/ROCK was detected by q RT-PCR ane western blotting after transfected with siRNA.TM cells were treated with ROCK inhibitors for 24 hours,and the activity of the Wnt/β-catenin signaling pathway were detected by q RT-PCR,western blotting,luciferase reporter and TOPFlash reporter assays.Results:1.The mRNA level of SNHG11 was downregulated in GTM₃ cells and the glaucoma mouse model compared with iHTM cells and control mouse.2.Knockdown of SNHG11 inhibited the survivability and migration of TM cells,activating autophagy and apoptosis,indicating that SNHG11 may participate in the pathogenesis of glaucoma by affecting the proliferation,migration,autophagy,apoptosis,and other functions of its cells.3.The Wnt/β-catenin signalling pathway was repressed in GTM₃ cells,and the glaucoma mouse model compared with iHTM cells and control mouse.4.After knocking down of lncRNA SNHG11,the Wnt/β-catenin signaling pathway was also inhibited.Possible targets are phosphorylation of Ser675 phosphorylation sites ofβ-catenin or phosphorylation of Ser33/37/Thr41 sites associated with GSK-3β.5.After using Wnt pathway inhibitors or activators and transfected with siRNA,the Wnt/β-catenin signaling pathway was inhibited,and the same is true for its targets as described above.6.In TM cells,Rho/ROCK is activated after lncRNA SNHG11 was knockdown,Wnt//β-catenin signaling pathway activity was activated after treatment with ROCKi,suggesting that SNHG11 may regulate the Wnt pathway through Rho/ROCK,and its possible action site is the phosphorylation of Ser675 phosphorylation site ofβ-catenin or the phosphorylation of Ser33/37/Thr41 site associated with GSK-3β.Conclusion:1.lncRNA SNHG11 expression is downregulated in glaucoma,and SNHG11is involved in the survival,migration,apoptosis,and autophagy of TM cells.2.Wnt/β-catenin signaling pathway is inhibited in glaucoma.3.lncRNA SNHG11 is an important regulator of the pathogenesis of glaucoma,mainly through Rho/ROCK regulation of Wnt/β-catenin signaling pathway affects the migration,apoptosis,autophagy and proliferation of TM cells,the possible sites are Ser675 phosphorylation site ofβ-catenin and Ser33/37/Thr41 phosphorylation site associated with GSK-3β.4.lncRNA SNHG11 is involved in the pathogenesis of glaucoma and may be used as a future therapeutic target for glaucoma.