The effects of c-Src kinase on EMT signaling pathway in human lens epithelial cells associated with lens diseases

Abstract

Background: The signaling pathway of epithelial to mesenchymal transition (EMT) is regulated by c-Src kinase in many cells. The purpose of this study was to investigate the effects of c-Src kinase on EMT of human lens epithelial cells in vivo stimulated by different factors. Methods: Human lens epithelial cells, HLE-B3, were exposed to either an inflammatory factor, specifically IL-1α, IL-6, TNF-α or IL-1β, at 10 ng/mL or high glucose (35.5 mM) for 30 mins. Activity of c-Src kinase was evaluated by the expression of p-Src 418 with western blot assay. To investigate the effects of activation of c-Src on EMT, HLE-B3 cells were transfected with pCDNA3.1-Src Y530F to upregulate activity of c-Src kinase, and pSlience4.1-ShSrc to knock it down. The expressions of c-Src kinase and molecular markers of EMT such as E-cadherin, ZO-1, α-SMA, and Vimentin were examined at 48 h by RT-PCR and western blot. At 48 h and 72 h of transfection, cell proliferation was detected by MTT, and cell mobility and migration were determined by scratch and transwell assays. Results: Activity of c-Src kinase, which causes the expression of p-Src 418 , was upregulated by different inflammatory factors and high glucose in HLE-B3 cells. When HLE-B3 cells were transfected with pCDNA3.1-Src Y530F , the expression of c-Src kinase was upregulated on both mRNA and protein levels, and activity of c-Src kinase, expression of p-Src 418 increased. The expressions of both E-cadherin and ZO-1 were suppressed, while the expressions of vimentin and α-SMA were elevated on both mRNA and protein levels at the same time. Cell proliferation, mobility and migration increased along with activation of c-Src kinase. Conversely, when HLE-B3 cells were transfected with pSlience4.1-ShSrc, both c-Src kinase and p-Src 418 expressions were knocked down. The expressions of E-cadherin and ZO-1 increased, but the expressions of Vimentin and α-SMA decreased; meanwhile, cell proliferation, mobility and migration reduced. Conclusions: The c-Src kinase in lens epithelial cells is easily activated by external stimuli, resulting in the induction of cell proliferation, mobility, migration and EMT.

Diosmetin‐7‐O‐β‐D‐glucopyranoside suppresses endothelial–mesenchymal transformation through endoplasmic reticulum stress in cardiac fibrosis

“…Previous studies described the importance of the Src kinase pathway for EMT in colorectal cancer 33 and lens diseases; 34 Src phosphorylation was the assay indicator. Therefore, we examined the phosphorylation level of Src in CMECs.…”

“…16 Previous studies have shown that c-Src participates in EMT/EndMT of multiple cells. 16,[32][33][34] It is well known that GRP78 is existed in ER, which binds with PERK, IRE1 and ATF6. This comprehensive ER stress inhibition effect is also the advantage of natural active ingredients over specific small molecule inhibitors because even small molecule inhibitors have strong inhibition effect on a specific protein, UPR reaction can be activated by other ways.…”

Long-term myofibroblast persistence in the capsular bag contributes to the late spontaneous in-the-bag intraocular lens dislocation

“…VIM is also present in mesenchymal cells and different stages of cell development 93 . In vitro and in vivo criteria for type II EMT transition to fibroblasts include increased expression of VIM 94 , among others, and such changes have been studied in lens epithelium 95,96 . In vitro models for PCO have found LECs capable of proliferating and migrating on the IOL-LC complex's posterior capsule to be rich in VIM and actin 11 .…”

The Interaction Between Autophagy and Epithelial-Mesenchymal Transition via NICD/ULK1 is Involved in the Formation of Diabetic Cataracts

“…Epithelial-mesenchymal transition (EMT) involves loss of the epithelial characteristics of epithelial cells and acquisition of mesenchymal characteristics under various external stimuli (Dongre et al 2019). Various studies have shown that LECs undergo EMT under high glucose conditions in vitro or in cataracts of individuals with diabetic mellitus Li et al 2019; Han et al 2018;Du et al 2017). EMT activation leads to disruption of cell-cell junctions and cell polarity, degradation of the basement membrane, and extracellular matrix reorganization, manifested by decreased expression of epithelial markers such as E-cadherin and some kinds of cytokeratin and increased expression of mesenchymal markers such as α-SMA, N-cadherin, vimentin and others (Dongre et al 2019).…”