scholarly journals Long Non-Coding RNA H19 Regulates Human Lens Epithelial Cells Function

2018 ◽  
Vol 50 (1) ◽  
pp. 246-260 ◽  
Author(s):  
Xin Liu ◽  
Chang Liu ◽  
Kun Shan ◽  
Shujie Zhang ◽  
Yi Lu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Differentially Expressed ◽  
Human Lens ◽  
Lens Epithelial Cells ◽  
Double Staining ◽  
Human Lens Epithelial Cells ◽  
And Migration ◽  
Proliferation And Migration

Background/Aims: Age-related cataract (ARC) remains the leading cause of visual impairment among the elderly population. Long non-coding RNAs (lncRNAs) have emerged as potential regulators in many ocular diseases. However, the role of lncRNAs in nuclear ARC, a subtype of ARC, requires further elucidation. Methods: LncRNA sequencing was performed to identify differentially expressed lncRNAs between the capsules of transparent and nuclear ARC lenses. Expression validation was confirmed by qRT-PCR. MTT assay, Calcein-AM and propidium iodide double staining, Rhodamine 123 and Hoechst double staining, EdU and transwell assay were used to determine the role of H19 or miR-675 in the viability, apoptosis, proliferation and migration of primary cultured human lens epithelial cells (HLECs). Bioinformatics and luciferase reporter assays were used to identify the binding target of miR-675. Results: Sixty-three lncRNAs are differentially expressed between the capsules of transparent and nuclear ARC lenses. One top abundantly expressed lncRNA, H19, is significantly up-regulated in the nuclear ARC lens capsules and positively associated with nuclear ARC grade. H19 knockdown accelerates apoptosis development and reduces the proliferation and migration of HLECs upon oxidative stress. H19 is the precursor of miR-675, and a reduction of H19 inhibits miR-675 expression. miR-675 regulates CRYAA expression by targeting the binding site within the 3’UTR. Moreover, miR-675 increases the proliferation and migration while decreasing the apoptosis of HLECs upon oxidative stress. Conclusion: H19 regulates HLECs function through miR-675-mediated CRYAA expression. This finding would provide a novel insight into the pathogenesis of nuclear ARC.

scholarly journals Thioredoxin Binding Protein-2 Regulates Autophagy of Human Lens Epithelial Cells under Oxidative Stress via Inhibition of Akt Phosphorylation

2016 ◽  
Vol 2016 ◽  
pp. 1-17 ◽  
Author(s):  
Jiaojie Zhou ◽  
Ke Yao ◽  
Yidong Zhang ◽  
Guangdi Chen ◽  
Kairan Lai ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Cell Viability ◽  
Binding Protein ◽  
Negative Regulator ◽  
Human Lens ◽  
Lens Epithelial Cells ◽  
Human Lens Epithelial Cells ◽  
Age Related

Oxidative stress plays an essential role in the development of age-related cataract. Thioredoxin binding protein-2 (TBP-2) is a negative regulator of thioredoxin (Trx), which deteriorates cellular antioxidant system. Our study focused on the autophagy-regulating effect of TBP-2 under oxidative stress in human lens epithelial cells (LECs). Human lens epithelial cells were used for cell culture and treatment. Lentiviral-based transfection system was used for overexpression of TBP-2. Cytotoxicity assay, western blot analysis, GFP/mCherry-fused LC3 plasmid, immunofluorescence, and transmission electronic microscopy were performed. The results showed that autophagic response of LECs with increased LC3-II, p62, and GFP/mCherry-LC3 puncta (P<0.01) was induced by oxidative stress. Overexpression of TBP-2 further strengthens this response and worsens the cell viability (P<0.01). Knockdown of TBP-2 attenuates the autophagic response and cell viability loss induced by oxidative stress. TBP-2 mainly regulates autophagy in the initiation stage, which is mTOR-independent and probably caused by the dephosphorylation of Akt under oxidative stress. These findings suggest a novel role of TBP-2 in human LECs under oxidative stress. Oxidative stress can cause cell injury and autophagy in LECs, and TBP-2 regulates this response. Hence, this study provides evidence regarding the role of TBP-2 in lens and the possible mechanism of cataract development.

scholarly journals SiRNA Targeting mTOR Effectively Prevents the Proliferation and Migration of Human Lens Epithelial Cells

PLoS ONE ◽  
2016 ◽  
Vol 11 (12) ◽  
pp. e0167349 ◽  
Author(s):  
Chunmei Zhang ◽  
Jingjing Liu ◽  
Na Jin ◽  
Guiming Zhang ◽  
Yahui Xi ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Human Lens ◽  
Lens Epithelial Cells ◽  
Human Lens Epithelial Cells ◽  
And Migration ◽  
Proliferation And Migration

Role of Aldehyde Dehydrogenase Isozymes in the Defense of Rat Lens and Human Lens Epithelial Cells against Oxidative Stress

2005 ◽  
Vol 46 (1) ◽  
pp. 259 ◽  
Author(s):  
Sanjeev Choudhary ◽  
Tianlin Xiao ◽  
Leoncio A. Vergara ◽  
Sanjay Srivastava ◽  
David Nees ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Aldehyde Dehydrogenase ◽  
Human Lens ◽  
Lens Epithelial Cells ◽  
Human Lens Epithelial Cells

scholarly journals LncRNA FEZF1-AS1 Promotes TGF-β2-Mediated Proliferation and Migration in Human Lens Epithelial Cells SRA01/04

2019 ◽  
Vol 2019 ◽  
pp. 1-10
Author(s):  
Yong Wang ◽  
Lili Chen ◽  
Yonghui Gu ◽  
Ying Wang ◽  
You Yuan ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Antisense Rna ◽  
Noncoding Rna ◽  
Human Lens ◽  
Lens Epithelial Cells ◽  
Protein Levels ◽  
Human Lens Epithelial Cells ◽  
Cell Proliferation And Migration ◽  
And Migration ◽  
Proliferation And Migration

Posterior capsule opacification (PCO) is a common complication after cataract surgery attributed to the proliferation and migration of postoperative residual lens epithelial cells (LECs). The long noncoding RNA (lncRNA) FEZ family zinc finger 1 antisense RNA 1 (FEZF1-AS1) promotes the proliferation and migration of multiple types of cancer cells. Here, we discovered that FEZF1-AS1 is markedly upregulated in TGF-β2-treated SRA01/04 cells. In addition, the proliferation and migration of SRA01/04 cells were enhanced following TGF-β2 treatment. FEZF1-AS1 knockdown inhibited the TGF-β2-induced proliferation and migration of SRA01/04 cells. Accordingly, FEZF1-AS1 overexpression promoted the TGF-β2-induced proliferation and migration of SRA01/04 cells. Finally, FEZF1-AS1 upregulated TGF-β2-induced SRA01/04 cell proliferation and migration via boosting FEZF1 protein levels. Our findings indicate that the dysregulation of FEZF1-AS1 participates in the TGF-β2-induced proliferation and migration of human lens epithelial cells (HLECs), which might be achieved, at least in part, through the induction of FEZF1 expression.

scholarly journals The role of Trx/TBP-2 in oxidative stress induced autophagy in human lens epithelial cells

2019 ◽  
Author(s):  
Jianghua Hu ◽  
Lifang Shen ◽  
Chengshou Zhang ◽  
Ke Yao ◽  
Yibo Yu
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Cellular Response ◽  
Negative Regulator ◽  
Cell Counting ◽  
Human Lens ◽  
Lens Epithelial Cells ◽  
Mrna Levels ◽  
Human Lens Epithelial Cells

Abstract Background: Oxidative stress plays an important role in age-related cataract development. The cellular antioxidant protein thioredoxin (Trx) and its negative regulator, thioredoxin binding protein-2 (TBP-2), maintain the intracellular redox balance upon oxidative stress. The aim of this study is to investigate role of Trx and TBP-2 in human lens epithelial cells (LECs) under oxidative stress. Methods: LECs were treated with 50 μM of H2O2 serum-free medium for different duration, and the mRNA and protein levels of Trx-1, Trx-2 and TBP-2 were measured by reverse transcription-polymerase chain reaction and Western blot. Trx-1 activity was evaluated by Thioredoxin Activity Fluorescent Assay. The subcellular localization of Trx-1, Trx-2 and TBP-2 was evaluated by cellular immunofluorescence. The cell viability was detected by Cell Counting Kit-8 (CCK-8) and the LC3-II protein level was detected to evaluate the autophagy level. The interaction between Trx-2 and TBP-2 was examined by Co-Immunoprecipitation (Co-IP). Results: The results showed that the mRNA levels of the Trx-1, Trx-2 and TBP-2 were kinetically changed after treatment with 50 μM of H2O2 for different duration. Exposure to H2O2 increased the expression of Trx-2 and TBP-2 but not Trx-1, while the exposure inhibited Trx-1 activity. TBP-2 was co-localized with Trx-1 and Trx-2, exposure to H2O2 enriched co-localization of TBP-2 to Trx-1 but not Trx-2, and increased the interaction between TBP-2 and Trx-1. Under normal circumstances, Trx-1 over-expression enhanced autophagic response and mainly regulates autophagy in the initiation. Conclusions: This study demonstrates differential roles of Trx-1 and Trx-2 in cellular response to oxidative stress, and oxidative stress increased Trx-1 interacting to TBP-2 and Trx-1 regulating autophagic response.

Mitochondria-Targeted Antioxidant Peptide SS31 Protects Cultured Human Lens Epithelial Cells against Oxidative Stress

2014 ◽  
Vol 40 (8) ◽  
pp. 822-829 ◽  
Author(s):  
Meng Cai ◽  
Jing Li ◽  
Shaofen Lin ◽  
Xiaoyun Chen ◽  
Juan Huang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Human Lens ◽  
Lens Epithelial Cells ◽  
Antioxidant Peptide ◽  
Human Lens Epithelial Cells

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

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Xingyu Li ◽  
Fang Wang ◽  
Meixia Ren ◽  
Minjuan Du ◽  
Jian Zhou
Keyword(s):  
Cell Proliferation ◽  
Epithelial Cells ◽  
Western Blot ◽  
Src Kinase ◽  
Human Lens ◽  
Lens Epithelial Cells ◽  
Protein Levels ◽  
Human Lens Epithelial Cells ◽  
And Migration ◽  
E Cadherin

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-Src418 with western blot assay. To investigate the effects of activation of c-Src on EMT, HLE-B3 cells were transfected with pCDNA3.1-SrcY530F 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-Src418, was upregulated by different inflammatory factors and high glucose in HLE-B3 cells. When HLE-B3 cells were transfected with pCDNA3.1-SrcY530F, the expression of c-Src kinase was upregulated on both mRNA and protein levels, and activity of c-Src kinase, expression of p-Src418 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-Src418 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.

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