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Updated: Oct 19, 2025
Molecular Analysis of Endothelial-mesenchymal Transition Induced by Transforming Growth Factor-β Signaling
Published on: August 3, 2018
miRNA-26b suppresses the TGF-β2-induced progression of HLE-B3 cells via the PI3K/Akt pathway
En Shi1, Xiang-Nan Ye2, Liu-Yi Xie3
1Department of Ophthalmology, Ningbo Eye Hospital, Ningbo 315040, Zhejiang Province, China.
Aim:
To study the effect of miR-26b on lens epithelial cells induced by transforming growth factor beta (TGF-β) 2 and the underlying signaling pathways.
Methods:
Human lens epithelial cell line B-3 (HLE-B3) was incubated with TGF-β2 (5 ng/mL) and then transfected with miR-26b mimics. The expression of miR-26b was determined using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), while 5'-bromodeoxyuridine (BrdU) and wound-healing assays were used to measure the growth and migration of HLE-B3 cells, respectively. The expression of epithelial-mesenchymal transition (EMT) markers and the activity of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway were measured by Western blotting assay and immunofluorescence staining. Electron microscopy was also used to observe cellular morphology.
Results:
The expression levels of miR-26b were significantly reduced in human posterior capsular opacification-attached lens tissue and TGF-β2-stimulated HLE-B3 cells. In the presence of TGF-β2, the growth, migration, and EMT of HLE-B3 cells were distinctly enhanced; these effects were attenuated by the administration of miR-26b mimics. Furthermore, the overexpression of miR-26b significantly reduced upregulation of the PI3K/Akt pathway when stimulated by TGF-β2 in HLE-B3 cells. Moreover, the addition of an activator (740 Y-P) led to the upregulation of the PI3K/Akt pathway and abolished the protective effect of miR-26b on the HLE-B3 cells that was mediated by TGF-β2.
Conclusion:
The miR-26b suppresses TGF-β2-induced growth, migration, and EMT in HLE-B3 cells by regulating the PI3K/Akt signaling pathway.
Insights
MicroRNA-26b (miR-26b) inhibits transforming growth factor beta 2 (TGF-β2)-induced cell changes in lens epithelial cells. It suppresses cell growth, migration, and epithelial-mesenchymal transition by regulating the PI3K/Akt pathway.
Area of Science:
- Ophthalmology
- Molecular Biology
- Cell Biology
Background:
- Transforming growth factor beta 2 (TGF-β2) is implicated in lens epithelial cell dysfunction.
- MicroRNAs (miRNAs) play crucial roles in regulating cellular processes.
- Understanding miRNA involvement in TGF-β2-induced cellular changes is vital for eye research.
Purpose of the Study:
- To investigate the role of miR-26b in TGF-β2-induced lens epithelial cells.
- To elucidate the underlying signaling pathways affected by miR-26b and TGF-β2.
Main Methods:
- Human lens epithelial cells (HLE-B3) were treated with TGF-β2 and transfected with miR-26b mimics.
- Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) assessed miR-26b expression.
- Cell growth, migration, epithelial-mesenchymal transition (EMT), and PI3K/Akt pathway activity were evaluated using various assays including BrdU, wound-healing, Western blotting, and immunofluorescence.
Main Results:
- miR-26b expression was reduced in posterior capsular opacification and TGF-β2-treated cells.
- TGF-β2 enhanced HLE-B3 cell growth, migration, and EMT, effects attenuated by miR-26b mimics.
- miR-26b overexpression suppressed TGF-β2-induced PI3K/Akt pathway activation, which was reversed by a PI3K/Akt activator.
Conclusions:
- miR-26b acts as a suppressor of TGF-β2-induced cellular responses in lens epithelial cells.
- The PI3K/Akt signaling pathway is a key mediator in the miR-26b- TGF-β2 interaction.
- Targeting miR-26b may offer therapeutic potential for conditions involving TGF-β2-induced lens epithelial cell changes.
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