Silencing of ILK attenuates the abnormal proliferation and migration of human Tenon's capsule fibroblasts induced by TGF-β2.
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.
Medical School of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
Yao
Y
Xing
Department of Ophthalmology, The First Affiliated Hospital of Medical School of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
Lijun
L
Cui
Department of Ophthalmology, The First Affiliated Hospital of Medical School of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
Qianyan
Q
Kang
D000199:Actins / Q000378:metabolism; D015703:Antigens, CD; D015153:Blotting, Western; D015820:Cadherins / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D019938:Cyclin D1 / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology*; D005455:Fluorescent Antibody Technique; D059585:G1 Phase Cell Cycle Checkpoints / Q000187:drug effects; D020868:Gene Silencing / Q000187:drug effects*; D006801:Humans; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D058475:Tenon Capsule / Q000473:pathology*; D053781:Transforming Growth Factor beta2 / Q000494:pharmacology*; D000099200:Scaffold Protein ILK
D000199:Actins; D015703:Antigens, CD; D015820:Cadherins; D019938:Cyclin D1; D017346:Protein Serine-Threonine Kinases; D012333:RNA, Messenger; D053781:Transforming Growth Factor beta2; D000099200:Scaffold Protein ILK; C541116:ACTA2 protein, human; C513384:CDH1 protein, human