Dataset Information

Aldose Reductase Mediates Transforming Growth Factor ?2 (TGF-?2)-Induced Migration and Epithelial-To-Mesenchymal Transition of Lens-Derived Epithelial Cells.

ABSTRACT: Cataract surgery involves removal of lens tissue, but is associated with a high complication rate due to regrowth of residual lens epithelial cells to produce posterior capsule opacification (PCO) and diminished visual acuity. As inhibitors of aldose reductase (AR) have been shown to suppress markers of PCO, our studies were designed to identify a role for AR in the pathogenesis of PCO.Sorbinil-mediated AR inhibition was determined by measuring sorbitol accumulation. Cell migration was measured using both transwell and scratch assays. Proteins in the SMAD signaling pathway were measured by Western blotting. The interactions of AR and SMADs were demonstrated by co-immunoprecipitation (Co-IP) and proximity ligation assay (PLA). Epithelial-to-mesenchymal transition (EMT) expression was measured by Western blot and quantitative PCR (q-PCR). Matrix metalloproteinase (MMP)-2 and MMP-9 activities were measured in conditioned medium by zymography.We observed that either Sorbinil-mediated AR inhibition or siRNA-mediated AR gene knockdown prevented migration of lens epithelial cells following exposure to TGF-?2. AR inhibition or AR knockdown reduced SMAD and MMP activation triggered by TGF-?2. In addition, we demonstrated AR inhibition or AR knockdown decreased TGF-?2-induced expression of EMT markers. Co-IP studies and PLA were used to demonstrate that AR and SMAD2 interact either directly or in close concert with additional factor(s) in a nonenzymatic manner.This study demonstrates that AR participates in the response of lens epithelial cells to TGF-?2. Our studies raise the possibility that AR inhibition may be effective in preventing development of PCO by disrupting the TGF-?2/SMAD pathway.

SUBMITTER: Chang KC

PROVIDER: S-EPMC4495811 | biostudies-other | 2015 Jul

REPOSITORIES: biostudies-other

ACCESS DATA

Publications

Aldose Reductase Mediates Transforming Growth Factor β2 (TGF-β2)-Induced Migration and Epithelial-To-Mesenchymal Transition of Lens-Derived Epithelial Cells.

Chang Kun-Che KC Petrash J Mark JM

Investigative ophthalmology & visual science 20150701 8

<h4>Purpose</h4>Cataract surgery involves removal of lens tissue, but is associated with a high complication rate due to regrowth of residual lens epithelial cells to produce posterior capsule opacification (PCO) and diminished visual acuity. As inhibitors of aldose reductase (AR) have been shown to suppress markers of PCO, our studies were designed to identify a role for AR in the pathogenesis of PCO.<h4>Methods</h4>Sorbinil-mediated AR inhibition was determined by measuring sorbitol accumulati ...[more]

PMID: 26132779

Similar Datasets

Project description:A growing body of evidence supports that the epithelial-to-mesenchymal transition (EMT), which occurs during cancer development and progression, has a crucial role in metastasis by enhancing the motility of tumor cells. Transforming growth factor-? (TGF-?) is known to induce EMT in a number of cancer cell types; however, the mechanism underlying this transition process is not fully understood. In this study we have demonstrated that TGF-? upregulates the expression of tumor suppressor protein Par-4 (prostate apoptosis response-4) concomitant with the induction of EMT. Mechanistic investigations revealed that exogenous treatment with each TGF-? isoform upregulates Par-4 mRNA and protein levels in parallel levels of phosphorylated Smad2 and I?B-? increase. Disruption of TGF-? signaling by using ALK5 inhibitor, neutralizing TGF-? antibody or phosphoinositide 3-kinase inhibitor reduces endogenous Par-4 levels, suggesting that both Smad and NF-?B pathways are involved in TGF-?-mediated Par-4 upregulation. NF-?B-binding sites in Par-4 promoter have previously been reported; however, using chromatin immunoprecipitation assay we showed that Par-4 promoter region also contains Smad4-binding site. Furthermore, TGF-? promotes nuclear localization of Par-4. Prolonged TGF-?3 treatment disrupts epithelial cell morphology, promotes cell motility and induces upregulation of Snail, vimentin, zinc-finger E-box binding homeobox 1 and N-Cadherin and downregulation of Claudin-1 and E-Cadherin. Forced expression of Par-4, results in the upregulation of vimentin and Snail expression together with increase in cell migration. In contrast, small interfering RNA-mediated silencing of Par-4 expression results in decrease of vimentin and Snail expression and prevents TGF-?-induced EMT. We have also uncovered a role of X-linked inhibitor of apoptosis protein in the regulation of endogenous Par-4 levels through inhibition of caspase-mediated cleavage. In conclusion, our findings suggest that Par-4 is a novel and essential downstream target of TGF-? signaling and acts as an important factor during TGF-?-induced EMT.

Dataset Information

Aldose Reductase Mediates Transforming Growth Factor ?2 (TGF-?2)-Induced Migration and Epithelial-To-Mesenchymal Transition of Lens-Derived Epithelial Cells.

Publications

Aldose Reductase Mediates Transforming Growth Factor β2 (TGF-β2)-Induced Migration and Epithelial-To-Mesenchymal Transition of Lens-Derived Epithelial Cells.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure