Authors: Liu Aihua, Gao Meizi, Huang Liangyu, Liu Xun, Su Ruihong, Zhao Jinzhi, Wang Liming, Zhang Xiaomin, Li Xiaorong, Dong Lijie
Abstract [View PDF] [Read Full Text]
Objective
To explore the role of forkhead box F2 (FoxF2) in the extracellular matrix of trabecular meshwork.
Methods
The cultured human trabecular meshwork cells (HTMCs) were divided into Scramble control group and FoxF2 small hairpin RNA (shRNA) group, then FoxF2 shRNA, the FoxF2 restructuring interference carrier was built, HTMCs were infected with FoxF2 shRNA lentivirus.Western blot assay was used to detect the expression of FoxF2 protein and extracellular matrix.Furthermore, Transwell counting experiment was used to analyze the migration ability of HTMCs.
Results
The cultured HTMCs grew well and showed a long spindle shape.The growth status of HTMCs was well, and their morphological characteristics were consistent with the HTMCs in vivo.The relative expression level of FoxF2 protein in the FoxF2 shRNA group was lower than that in the Scramble control group, with a significant difference between them (0.72±0.02 vs.1.27±0.05; t=16.68, P<0.01). The relative expression level of fibronectin (FN), collagen typeⅠ(COLⅠ) and α-smooth muscle actin (α-SMA) were 0.43±0.03, 0.53±0.08 and 0.86±0.15 in the FoxF2 shRNA group, and 0.87±0.04, 1.66±0.06 and 1.73±0.13 in the Scramble control group, respectively, the relative expression levels of FN, COLⅠ and α-SMA in the FoxF2 shRNA group were significantly lower than those in the Scramble control group (t=15.08, 18.81, 7.50, all at P<0.01). The migration number of HTMCs in the FoxF2 shRNA group was significantly lower than that in the Scramble control group (117.30±11.41 vs. 251.00±10.37; t=8.72, P<0.01).
Conclusions
The FoxF2 shRNA lentivirus are successfully constructed, which can decrease the expression of FoxF2 in HTMCs.Low expression of FoxF2 can reduce the expression level of extracellular matrix protein in HTMCs and inhibit the migration ability of HTMCs.