Effects of Long-Chain Non-Coding Ribonucleic Acid Hox Tran Antisense RNA on Proliferation and Migration of Epithelial Cells of Cataract Lens

2021 ◽  
Vol 11 (12) ◽  
pp. 2329-2336
Author(s):  
Qiang Zeng ◽  
Yiting Luo

In order to explore effects of long-chain non-coding ribonucleic acid (RNA) HOTAIR on proliferation and migration of human lens epithelial cells, SRA01/04 cells were selected as the research strain in this study and divided into S1 group (no HOTAIR transfection), S2 group (siHOTAIR transfection), S3 group (siHOTAIR+10 ng/mL TGF-β2), and S4 group (no HOTAIR transfection+10 ng/mL TGF-β2) according to the presence or absence of transforming growth factor (TGF)-β2 and silent HOTAIR treatment. 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) colorimetric method was applied to detect cell proliferation.Western blot was used for detection of E-cadherin, zonula occluden-1 (ZO-1), Vimentin, α-smooth muscle actin (SMA), Snail, Slug, zinc finger E-box binding homeobox 1 (ZEB1), and Smad-2 expressions. Results showed that the number of transmembrane cells in S4 group was higher markedly than that of the other groups, but that of S2 group dropped steeply compared with the other groups (P <0.05); E-cadherin (2.59±0.58) and ZO-1 (1.95±0.56) of S2 group increased hugely compared with the other groups, while Vimentin (0.57±0.14) and α-SMA (0.64±0.28) decreased sharply compared with the other groups (P < 0.05); Snail (2.51±0.59), Slug (2.11±0.47), and ZEB1 (2.83±0.53) of S4 group rose obviously compared with the other groups, but the above of S2 group reduced hugely compared with the other groups (P < 0.05); pSmad-2 and pSmad-3 of S4 group elevated greatly compared with the other groups, and those of S2 group reduced hugely compared with the other groups (P < 0.05). In conclusion, HOTAIR high expression could promote TGF-β2-induced SRA01/04 cell proliferation, migration, invasion, and epithelial-mesenchymal trans-differentiation, which was related to TGF-β/Smad signaling pathway.

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Xingyu Li ◽  
Fang Wang ◽  
Meixia Ren ◽  
Minjuan Du ◽  
Jian Zhou

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.


2018 ◽  
Vol 50 (1) ◽  
pp. 246-260 ◽  
Author(s):  
Xin Liu ◽  
Chang Liu ◽  
Kun Shan ◽  
Shujie Zhang ◽  
Yi Lu ◽  
...  

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.


PLoS ONE ◽  
2016 ◽  
Vol 11 (12) ◽  
pp. e0167349 ◽  
Author(s):  
Chunmei Zhang ◽  
Jingjing Liu ◽  
Na Jin ◽  
Guiming Zhang ◽  
Yahui Xi ◽  
...  

2019 ◽  
Author(s):  
Xing-Yu Li ◽  
Fang Wang ◽  
Mei-Xia Ren ◽  
Min-Juan Du ◽  
Jian Zhou

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.


2019 ◽  
Author(s):  
Xing-Yu Li ◽  
Fang Wang ◽  
Mei-Xia Ren ◽  
Min-Juan Du ◽  
Jian Zhou

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.


2019 ◽  
Vol 2019 ◽  
pp. 1-10
Author(s):  
Yong Wang ◽  
Lili Chen ◽  
Yonghui Gu ◽  
Ying Wang ◽  
You Yuan ◽  
...  

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.


2019 ◽  
Author(s):  
Xing-Yu Li ◽  
Fang Wang ◽  
Mei-Xia Ren ◽  
Min-Juan Du ◽  
Jian Zhou

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.


Author(s):  
Jin-Soo Park ◽  
RyeonJin Cho ◽  
Eun-Young Kang ◽  
Yeon-Mok Oh

AbstractEmphysema, a pathological component of chronic obstructive pulmonary disease, causes irreversible damage to the lung. Previous studies have shown that Slit plays essential roles in cell proliferation, angiogenesis, and organ development. In this study, we evaluated the effect of Slit2 on the proliferation and migration of mouse lung epithelial cells and its role in regeneration in an emphysema lung mouse model. Here, we have shown that Slit2/Robo signaling contributes to the regeneration of lungs damaged by emphysema. Mouse epithelial lung cells treated with Slit2 exhibited increased proliferation and migration in vitro. Our results also showed that Slit2 administration improved alveolar regeneration in the emphysema mouse model in vivo. Furthermore, Slit2/Robo signaling increased the phosphorylation of ERK and Akt, which was mediated by Ras activity. These Slit2-mediated cellular signaling processes may be involved in the proliferation and migration of mouse lung epithelial cells and are also associated with the potential mechanism of lung regeneration. Our findings suggest that Slit2 administration may be beneficial for alveolar regeneration in lungs damaged by emphysema.


2021 ◽  
pp. 153537022110281
Author(s):  
Yu Hou ◽  
Yu-Xi He ◽  
Jia-Hao Zhang ◽  
Shu-Rong Wang ◽  
Yan Zhang

Epithelial tissue has important functions such as protection, secretion, and sensation. Epithelial damage is involved in various pathological processes. Bone morphogenetic proteins (BMPs) are a class of growth factors with multiple functions. They play important roles in epithelial cells, including in differentiation, proliferation, and migration during the repair of the epithelium. This article reviews the functions and mechanisms of the most profoundly studied BMPs in the process of epithelial damage repair and their clinical significance.


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