Characterisation of TGF-β2 signalling and function in a human lens cell line

Wormstone, I. Michael; Tamiya, Shigeo; Eldred, Julie A.; Lazaridis, Konstantinos; Chantry, Andrew; Reddan, John R.; Anderson, Ian K.; Duncan, G.

doi:10.1016/j.exer.2003.08.006

Cited by 77 publications

(83 citation statements)

References 16 publications

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“…Numerous studies have implicated the TGFβ/Smad signaling pathway in PCO (Saika et al ., 2004; Wormstone et al ., 2004). αSMA, MMP2, CTGF, integrin αV, integrin α5, integrin β1, miR4279, and miR1469 have been shown to be upregulated by TGFβ2 during the EMT of HLE cells (Dawes et al ., 2007; Wang et al ., 2013; Mamuya et al ., 2014).…”

Section: Resultsmentioning

confidence: 99%

AGEs in human lens capsule promote the TGFβ2‐mediated EMT of lens epithelial cells: implications for age‐associated fibrosis

et al. 2016

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SummaryProteins in basement membrane (BM) are long‐lived and accumulate chemical modifications during aging; advanced glycation endproduct (AGE) formation is one such modification. The human lens capsule is a BM secreted by lens epithelial cells. In this study, we have investigated the effect of aging and cataracts on the AGE levels in the human lens capsule and determined their role in the epithelial‐to‐mesenchymal transition (EMT) of lens epithelial cells. EMT occurs during posterior capsule opacification (PCO), also known as secondary cataract formation. We found age‐dependent increases in several AGEs and significantly higher levels in cataractous lens capsules than in normal lens capsules measured by LC‐MS/MS. The TGFβ2‐mediated upregulation of the mRNA levels (by qPCR) of EMT‐associated proteins was significantly enhanced in cells cultured on AGE‐modified BM and human lens capsule compared with those on unmodified proteins. Such responses were also observed for TGFβ1. In the human capsular bag model of PCO, the AGE content of the capsule proteins was correlated with the synthesis of TGFβ2‐mediated α‐smooth muscle actin (αSMA). Taken together, our data imply that AGEs in the lens capsule promote the TGFβ2‐mediated fibrosis of lens epithelial cells during PCO and suggest that AGEs in BMs could have a broader role in aging and diabetes‐associated fibrosis.

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Section: Resultsmentioning

confidence: 99%

AGEs in human lens capsule promote the TGFβ2‐mediated EMT of lens epithelial cells: implications for age‐associated fibrosis

et al. 2016

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“…FHL-124 is a non-virally transformed cell line generated from human capsuleepithelial explants (Reddan et al, 1999) showing a 99.5% homology (in transcript profile) with the native lens epithelium (Wormstone et al, 2004). FHL-124 were routinely cultured at 35 C in a humidified atmosphere of 95% air and 5% CO 2 , in Eagle's Minimal Essential Medium (EMEM) (Gibco, Paisley, UK) supplemented with 5% v/v foetal bovine serum (Gibco) and 0.005% w/v gentamicin.…”

Section: Cell Culturementioning

confidence: 99%

Hypoxia-inducible factor-1 (HIF-1) pathway activation by quercetin in human lens epithelial cells

Radreau

Rhodes

Mithen

et al. 2009

Experimental Eye Research

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Keywords: quercetin flavonoid antioxidant diet lens HIF-1a hypoxia VEGF a b s t r a c t Quercetin is a dietary bioflavonoid which has been shown to inhibit lens opacification in a number of models of cataract. The objectives of this study were to determine gene expression changes in human lens epithelial cells in response to quercetin and to investigate in detail the mechanisms underlying the responses. FHL-124 cells were treated with quercetin (10 mM) and changes in gene expression were measured by microarray. It was found that 65% of the genes with increased expression were regulated by the hypoxia-inducible factor-1 (HIF-1) pathway. Quercetin (10 and 30 mM) induced a time-dependent increase in HIF-1a protein levels. Quercetin (30 mM) was also responsible for a rapid and long-lasting translocation of HIF-1a from the cytoplasm to the nucleus. Activation of HIF-1 signaling by quercetin was confirmed by qRT-PCR which showed upregulation of the HIF-1 regulated genes EPO, VEGF, PGK1 and BNIP3. Analysis of medium taken from FHL-124 cells showed a sustained dose-dependent increase in VEGF secretion following quercetin treatment. The quercetin-induced increase and nuclear translocation of HIF-1a was reversed by addition of excess iron (100 mM). These results demonstrate that quercetin activates the HIF-1 signaling pathway in human lens epithelial cells.

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“…The ability to monitor changes within a well-defined system is critical to understand the nature of both the induction of a wound healing response and fibrotic changes. Models available to study fibrosis such as cell lines [12], transgenic models [13,14] and tissue culture [15 -17] are well characterized in the lens. Postmortem analysis and clinical data also greatly aid the understanding of fibrosis in the lens [17 -19]; this information may not strictly inform us of the causal factors, but importantly the data do serve as a blueprint of changes that are seen in patients.…”

Section: The Lens: a Model For Fibrosismentioning

confidence: 99%

“…A large complement of methods is available to study PCO; cell lines provide an abundant and reliable source of material to investigate the relationship between signalling pathways and functional effects including cell growth, transdifferentiation, matrix deposition and contraction [5,12,30,31]. Tissue culture models have also served the research field well, and include lens explants [16] and in vitro capsular bag models [17,32 -34].…”

Section: The Lens: a Model For Fibrosismentioning

confidence: 99%

The lens as a model for fibrotic disease

Eldred

Dawes

Wormstone

2011

Phil. Trans. R. Soc. B

106

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Fibrosis affects multiple organs and is associated with hyperproliferation, cell transdifferentiation, matrix modification and contraction. It is therefore essential to discover the key drivers of fibrotic events, which in turn will facilitate the development of appropriate therapeutic strategies. The lens is an elegant experimental model to study the processes that give rise to fibrosis. The molecular and cellular organization of the lens is well defined and consequently modifications associated with fibrosis can be clearly assessed. Moreover, the avascular and non-innervated properties of the lens allow effective in vitro studies to be employed that complement in vivo systems and relate to clinical data. Using the lens as a model for fibrosis has direct relevance to millions affected by lens disorders, but also serves as a valuable experimental tool to understand fibrosis per se.

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Characterisation of TGF-β2 signalling and function in a human lens cell line

Cited by 77 publications

References 16 publications

AGEs in human lens capsule promote the TGFβ2‐mediated EMT of lens epithelial cells: implications for age‐associated fibrosis

AGEs in human lens capsule promote the TGFβ2‐mediated EMT of lens epithelial cells: implications for age‐associated fibrosis

Hypoxia-inducible factor-1 (HIF-1) pathway activation by quercetin in human lens epithelial cells

The lens as a model for fibrotic disease

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