Silencing of ILK attenuates the abnormal proliferation and migration of human Tenon's capsule fibroblasts induced by TGF-β2

Abstract

The cytokine, transforming growth factor-β (TGF-β), plays a key role in wound healing and tissue repair. Integrin-linked kinase (ILK) is a downstream factor of the TGF-β signaling pathway. Research on ILK has mainly focused on its role in the invasion and metastasis of cancer cells. However, little has been reported on the effects of ILK in human Tenon's capsule fibroblasts (HTFs). In this study, we investigated the role of ILK in the proliferation and migration of HTFs exposed to TGF-β2. A lentiviral vector targeting ILK was screened from three candidates and the experimental result indicated that RNA interference can be used to inhibit ILK expression at both the mRNA and protein level. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to assess ILK mRNA expression. Cell proliferation was quantified by MTT assay and cell cycle progression was detected by flow cytometric analysis. Migration was measured by wound healing assay. It was observed that the silencing of ILK suppressed the TGF-β2-induced proliferation of HTFs and led to G1 phase cell cycle arrest and the significant downregulation of cyclin D1 expression. The migration ability of the HTFs decreased following the silencing of ILK, while the downregulation of α-smooth muscle actin expression and the upregulation of E-cadherin expression were observed. The findings of our study indicate that the silencing of ILK attenuates the abnormal proliferation and migration of HTFs induced by TGF-β2, which reveals the therapeutic potential of ILK inhibition in the prevention of scarring following glaucoma filtration surgery.

Anti-Metastatic Effects of Plant Sap-Derived Extracellular Vesicles in a 3D Microfluidic Cancer Metastasis Model

“…Blocking PDGFC has been reported to reduce the growth and angiogenesis of tumors that are resistant to anti-VEGF therapies [ 32 , 33 , 34 ]. The ILK gene, a downstream target of TFG-β, was also reduced by DMS-EVs ( Figure 5 b); interestingly, studies have shown that silencing ILK inhibits cancer cells stimulated by TFG-β2 [ 35 ]. AKT1 was also downregulated by 1 and 10 µg/mL of DMS-EVs ( Figure 5 b).…”

“…CD44, a cell surface adhesion receptor that mediates cell–ECM interactions, can activate various signaling pathways that lead to cell proliferation, migration, and invasion. Studies have shown that knocking out CD44 suppresses the EMT phenotype ( Figure 5 c) [ 35 ].…”

<p>Emodin Reverses the Epithelial–Mesenchymal Transition of Human Endometrial Stromal Cells by Inhibiting ILK/GSK-3β Pathway</p>

“…30 Additionally, knockout of ILK decreased the invasion and metastasis abilities of cells through inhibiting the EMT process. 31 , 32 Considering the crucial roles of EMT in modulating cell migration and invasion, ILK may be a promising treatment target for various diseases. Tang et al discovered that emodin inhibited the expression of ILK through the crosstalk of AMPKα and ERK1/2 signaling.…”

MicroR-542-3p can mediate ILK and further inhibit cell proliferation, migration and invasion in osteosarcoma cells

“…Integrin-linked kinase ( ILK ) is a serine/threonine protein kinase located in focal adhesions, which promotes cell growth, proliferation, differentiation, migration and invasion [15]. ILK expression and activity have been revealed to be increased in association with the tumor grade, T status, lymph node metastasis and survival rate of lung cancer patients [16].…”

“…The overexpression of ILK has also been discovered to facilitate glioma cell invasion and migration and down-regulate E-cadherin. Besides, the inhibition of ILK has been shown to lead to cell cycle stagnation and stimulate apoptosis in PTEN-negative prostate cancer cells [15]. Indeed, previous studies have demonstrated that the increased ILK expression in poorly differentiated thyroid cancer and confirmed the relationship between ILK overexpression and poor prognosis [17].…”