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

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

Cellular Physiology and Biochemistry ◽

10.1159/000494003 ◽

2018 ◽

Vol 50 (1) ◽

pp. 246-260 ◽

Cited By ~ 6

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.

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

PLoS ONE ◽

10.1371/journal.pone.0167349 ◽

2016 ◽

Vol 11 (12) ◽

pp. e0167349 ◽

Cited By ~ 3

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

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

10.21203/rs.2.9375/v3 ◽

2019 ◽

Author(s):

Xing-Yu Li ◽

Fang Wang ◽

Mei-Xia Ren ◽

Min-Juan 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-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 hours by RT-PCR and western blot. At 48 hours and 72 hours of transfection, cell proliferation was detected by MTT, and cell mobility and migration were determined by scratch and transwell assay. 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 expression of both E-cadherin and ZO-1 was 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 expression were knocked down. The expression of E-cadherin and ZO-1 increased, but the expression 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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The effects of c-Src kinase on EMT signaling pathway in human lens epithelial cells associated with lens diseases

10.21203/rs.2.9375/v2 ◽

2019 ◽

Author(s):

Xing-Yu Li ◽

Fang Wang ◽

Mei-Xia Ren ◽

Min-Juan 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-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 hours by RT-PCR and western blot. At 48 hours and 72 hours of transfection, cell proliferation was detected by MTT, and cell mobility and migration were determined by scratch and transwell assay. 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 expression of both E-cadherin and ZO-1 was 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 expression were knocked down. The expression of E-cadherin and ZO-1 increased, but the expression 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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Effects of c-Src kinase on lens diseases associated with EMT of human lens epithelial cells

10.21203/rs.2.9375/v1 ◽

2019 ◽

Cited By ~ 1

Author(s):

Xing-Yu Li ◽

Fang Wang ◽

Mei-Xia Ren ◽

Min-Juan Du ◽

Jian Zhou

Keyword(s):

Cell Proliferation ◽

Epithelial Cells ◽

Western Blot ◽

Src Kinase ◽

Inflammatory Factors ◽

Human Lens ◽

Lens Epithelial Cells ◽

Human Lens Epithelial Cells ◽

And Migration ◽

E Cadherin

Abstract Background c-Src kinase regulated the signaling pathway of epithelial to mesenchymal transition (EMT) 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 exposure to inflammatory factors including IL-1α, IL-6, TNF-α, and IL-1β at 10 ng/mL, and high glucose (35.5 mM) respectively, for 30 mins. Activity of c-Src kinase was evaluated by expression of p-Src418 with western blot assay. To investigate activation of c-Src on EMT, HLE-B3 cells were transfected with pCDNA3.1-SrcY530F to up regulate c-Src, and pSlience4.1-ShSrc to knock down it. The expressions of c-Src kinase and molecular markers of EMT such as E-cadherin, ZO-1, α-SMA and vimentine were examined at 48 hours by RT-PCR and western blot. At 48 hours and 72 hours of transfection, cell proliferation was detected by MTT, cell mobility and migration were determined by scratch and transwell assay. Results Activity of c-Src kinase, 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 level, and activity of c-Src, p-Src418 increased. The expressions of E-cadherin and ZO-1 suppressed, while the expressions of vimentin and αSMA elevated on both mRNA and protein level 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 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 c-Src kinase in lens epithelial cells is easily activated by external stimuli, resulting in inducing of cell proliferation, mobility, migration and EMT.

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