The inhibitory effect of FoxF2 shRNA on the expression of extracellular matrix of human trabecular meshwork

Authors: Liu Aihua,  Gao Meizi,  Huang Liangyu,  Liu Xun,  Su Ruihong,  Zhao Jinzhi,  Wang Liming,  Zhang Xiaomin,  Li Xiaorong,  Dong Lijie

DOI: 10.3760/cma.j.issn.2095-0160.2019.06.002
Published 2019-06-10
Cite as Chin J Exp Ophthalmol, 2019,37(6): 405-410.

Abstract

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.

Key words:

Forkhead box F2; Trabecular meshwork cells; Extracellular matrix; Primary open angle glaucoma

Contributor Information

Liu Aihua
Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, School of Optometry and Ophthalmology, Tianjin Medical University, Tianjin 300384, China
Gao Meizi
Tianjin University of Traditional Chinese Medicine, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China
Huang Liangyu
Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, School of Optometry and Ophthalmology, Tianjin Medical University, Tianjin 300384, China
Liu Xun
Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, School of Optometry and Ophthalmology, Tianjin Medical University, Tianjin 300384, China
Su Ruihong
Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, School of Optometry and Ophthalmology, Tianjin Medical University, Tianjin 300384, China
Zhao Jinzhi
Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, School of Optometry and Ophthalmology, Tianjin Medical University, Tianjin 300384, China
Wang Liming
Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, School of Optometry and Ophthalmology, Tianjin Medical University, Tianjin 300384, China
Zhang Xiaomin
Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, School of Optometry and Ophthalmology, Tianjin Medical University, Tianjin 300384, China
Li Xiaorong
Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, School of Optometry and Ophthalmology, Tianjin Medical University, Tianjin 300384, China
Dong Lijie
Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, School of Optometry and Ophthalmology, Tianjin Medical University, Tianjin 300384, China
(Read 39 times, 1 visits today)
Updated: September 25, 2019 — 2:21 am