Dataset Information

Downregulation of transforming growth factor-? type II receptor prohibit epithelial-to-mesenchymal transition in lens epithelium.

ABSTRACT: PURPOSE: Transforming growth factor-? (TGF-?) is considered to be essential to induce epithelial-to-mesenchymal transition (EMT) which plays central roles in wound healing in ocular fibrotic complication. The present study investigates whether small interference RNAs (siRNAs) targeting the type II receptor of TGF-? (T?RII) could be used to minimize the TGF-? action. METHODS: TGF-? receptor type II (T?RII) specific siRNAs designed from the Nakamura human gene sequence were used to transfect the cultured lens epithelial cells (LECs). The optimal transfection of scramble siRNA-Cy3 labeled duplexes in cultured LECs were examined by laser scanning confocal microscope and flow cytometry. T?RII protein expression and transcript levels were analyzed by immunofluorescence, western blotting, and real time PCR, respectively. Western blotting was performed to examine protein expression of fibronectin and alpha-smooth muscle actin (?-SMA). Scratch assay was used to determine cell migration. Cell morphology was observed after transfection by inverted microscope. RESULTS: The optimal transfection rate of scramble siRNA-Cy3 labeled duplexes was efficient in that nearly to 50% in cultured LECs. T?RII specific siRNAs significantly reduced the receptor transcript and protein expression in cultured LECs. The gene knockdown inhibited LECs transdifferentiation, as it abrogated the expression of fibronectin and ?-SMA, and retarded cell migration on the scratch assay. In addition, after transfection with T?RII specific siRNA, the cultured LECs did not show fibroblast-like shape which was one of the feature signs of EMT. Wound scratch assays indicated that the number of cultured LECs migrated into the wounded area was significantly lower in T?RII specific siRNA treated group (12.8 ± 3.27/7.85 mm(2)), compared with normal (57.8 ± 3.06/7.85 mm(2)) and scrambled RNA transfected group (50.8 ± 3.64/7.85 mm(2); p<0.0001). CONCLUSIONS: Our results provided additional evidence to support that TGF-? pathway was involved in the development of EMT of human posterior capsule opacification, while how T?RII was involved should be further investigated.

SUBMITTER: Zheng D

PROVIDER: S-EPMC3365138 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Downregulation of transforming growth factor-β type II receptor prohibit epithelial-to-mesenchymal transition in lens epithelium.

Zheng Danying D Song Tingting T Zhongliu Xueying X Wu Mingxing M Liang Jingli J Liu Yizhi Y

Molecular vision 20120511

<h4>Purpose</h4>Transforming growth factor-β (TGF-β) is considered to be essential to induce epithelial-to-mesenchymal transition (EMT) which plays central roles in wound healing in ocular fibrotic complication. The present study investigates whether small interference RNAs (siRNAs) targeting the type II receptor of TGF-β (TβRII) could be used to minimize the TGF-β action.<h4>Methods</h4>TGF-β receptor type II (TβRII) specific siRNAs designed from the Nakamura human gene sequence were used to tr ...[more]

PMID: 22665970

Similar Datasets

Project description:Epithelial-mesenchymal transition (EMT) occurs in several disease states, including renal fibrosis and carcinogenesis. Myofibroblasts produced from EMT of renal tubular cells are responsible for the deposition of extracellular matrix components in a large portion of renal interstitial fibrosis. Transforming growth factor-beta (TGF-beta) plays an essential role in the EMT of renal tubular cells, but the molecular mechanism governing this process remains largely unknown. In this study, we found that RGC-32 (response gene to complement 32) is critical for TGF-beta-induced EMT of human renal proximal tubular cells (HPTCs). RGC-32 is not normally expressed in the HPTCs. However, TGF-beta stimulation markedly activates RGC-32 while inducing an EMT, as shown by the induction of smooth muscle alpha-actin (alpha-SMA) and extracellular matrix proteins collagen I and fibronectin, as well as the reduction of epithelial marker E-cadherin. TGF-beta function is mediated by several signaling pathways, but RGC-32 expression in HPTCs appears to be mainly regulated by Smad. Functionally, RGC-32 appears to mediate TGF-beta-induced EMT of HPTCs. Blockage of RGC-32 using short hairpin interfering RNA significantly inhibits TGF-beta induction of myofibroblast marker gene alpha-SMA while repressing the expression of E-cadherin. In contrast, overexpression of RGC-32 induces alpha-SMA expression while restoring E-cadherin. RGC-32 also inhibits the expression of another adherens junction protein, N-cadherin, suggesting that RGC-32 alone induces the phenotypic conversion of renal epithelial cells to myofibroblasts. Additional studies show that RGC-32 stimulates the production of extracellular matrix components fibronectin and collagen I. Mechanistically, RGC-32 induces EMT via the activation of other transcription factors such as Snail and Slug. RGC-32 knockdown inhibits the expression of Snail and Slug during TGF-beta-induced EMT. Taken together, our data demonstrate for the first time that RGC-32 plays a critical role in TGF-beta-induced EMT of renal tubular cells.

Dataset Information

Downregulation of transforming growth factor-? type II receptor prohibit epithelial-to-mesenchymal transition in lens epithelium.

Publications

Downregulation of transforming growth factor-β type II receptor prohibit epithelial-to-mesenchymal transition in lens epithelium.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure