AIM:To study the effect of mi R-26 b on lens epithelial cells induced by transforming growth factor beta(TGF-β)2 and the underlying signaling pathways.METHODS:Human lens epithelial cell line B-3(HLE-B3)was incubated ...AIM:To study the effect of mi R-26 b on lens epithelial cells induced by transforming growth factor beta(TGF-β)2 and the underlying signaling pathways.METHODS:Human lens epithelial cell line B-3(HLE-B3)was incubated with TGF-β2(5 ng/m L)and then transfected with mi R-26 b mimics.The expression of mi R-26 b was determined using quantitative reverse transcriptase polymerase chain reaction(q RT-PCR),while 5’-bromodeoxyuridine(Brd U)and wound-healing assays were used to measure the growth and migration of HLE-B3 cells,respectively.The expression of epithelialmesenchymal transition(EMT)markers and the activity of the phosphatidylinositol 3-kinase(PI3 K)/Akt pathway were measured by Western blotting assay and immunofluorescence staining.Electron microscopy was also used to observe cellular morphology.RESULTS:The expression levels of mi R-26 b were significantly reduced in human posterior capsular opacification-attached lens tissue and TGF-β2-stimulated HLE-B3 cells.In the presence of TGF-β2,the growth,migration,and EMT of HLE-B3 cells were distinctly enhanced;these effects were attenuated by the administration of mi R-26 b mimics.Furthermore,the overexpression of mi R-26 b significantly reduced upregulation of the PI3 K/Akt pathway when stimulated by TGF-β2 in HLE-B3 cells.Moreover,the addition of an activator(740 Y-P)led to the upregulation of the PI3 K/Akt pathway and abolished the protective effect of mi R-26 b on the HLE-B3 cells that was mediated by TGF-β2.CONCLUSION:The mi R-26 b suppresses TGF-β2-induced growth,migration,and EMT in HLE-B3 cells by regulating the PI3 K/Akt signaling pathway.展开更多
文摘AIM:To study the effect of mi R-26 b on lens epithelial cells induced by transforming growth factor beta(TGF-β)2 and the underlying signaling pathways.METHODS:Human lens epithelial cell line B-3(HLE-B3)was incubated with TGF-β2(5 ng/m L)and then transfected with mi R-26 b mimics.The expression of mi R-26 b was determined using quantitative reverse transcriptase polymerase chain reaction(q RT-PCR),while 5’-bromodeoxyuridine(Brd U)and wound-healing assays were used to measure the growth and migration of HLE-B3 cells,respectively.The expression of epithelialmesenchymal transition(EMT)markers and the activity of the phosphatidylinositol 3-kinase(PI3 K)/Akt pathway were measured by Western blotting assay and immunofluorescence staining.Electron microscopy was also used to observe cellular morphology.RESULTS:The expression levels of mi R-26 b were significantly reduced in human posterior capsular opacification-attached lens tissue and TGF-β2-stimulated HLE-B3 cells.In the presence of TGF-β2,the growth,migration,and EMT of HLE-B3 cells were distinctly enhanced;these effects were attenuated by the administration of mi R-26 b mimics.Furthermore,the overexpression of mi R-26 b significantly reduced upregulation of the PI3 K/Akt pathway when stimulated by TGF-β2 in HLE-B3 cells.Moreover,the addition of an activator(740 Y-P)led to the upregulation of the PI3 K/Akt pathway and abolished the protective effect of mi R-26 b on the HLE-B3 cells that was mediated by TGF-β2.CONCLUSION:The mi R-26 b suppresses TGF-β2-induced growth,migration,and EMT in HLE-B3 cells by regulating the PI3 K/Akt signaling pathway.
