Integrin β1-mediated signaling is involved in transforminggrowth factor-β2-promoted migration in human lens epithelial cells

[摘要] Purpose: Transforming growth factor-beta 2 (TGF-β2) is a potentgrowth inhibitor and apoptosis inducer. However, typically TGF-β2loses its growth-inhibitory and apoptosis-inducing effects butstimulates the migratory capacity of epithelial cells. In this study, weinvestigate the possible involvement of integrin and integrin-mediatedsignaling in TGF-β2-promoted migration and adhesion in human lensepithelial cells.Methods: A human lens epithelial cell line (HLE B-3) was treatedwith 100 pg/ml TGF-β2 for 6 h, 12 h, and 24 h. In vitro woundhealing assay and cell adhesion assay were performed to detect theeffect of TGF-β2 on HLEC adhesion and migration. Integrin β1expression changes in HLECs during the treatment of TGF-β2 weredetected in protein levels and mRNA levels using confocal microscopy,flow cytometric analysis, and real time quantitative reversetranscription polymerase chain reaction (RT-PCR). Focal adhesion kinase(FAK) activity was examined by FAK phosphorylation and total tyrosinephosphorylation during treatment with TGF-β2 in HLEC. Production ofendogenous TGF-β2 was measured by ELISA assay.Results: In this study, we found that TGF-β2 significantlystimulated cell adhesion and migration in HLECs. By immunofluorescencestaining and western blotting, we observed that TGF-β2 markedlyenhanced the expression of integrin β1 and the Tyr-phosphorylationof focal adhesion kinase (FAK). Real time quantitative RT-PCR alsoshowed the mRNA level of integrin β1 was upregulated. Neutralizinganti-integrin β1 monoclonal antibody significantly (p<0.05)inhibited TGF-β2-promoted HLEC adhesion and migration.Conclusions: TGF-β2 promoted HLEC adhesion and migration invitro. Integrin β1 and integrin-mediated signaling are necessaryfor TGF-β2-promoted adhesion and migration in human lens epithelialcells.

[学科分类] 生物化学/生物物理