Regulation of bFGF and TGF-β2 in human scleral fibroblasts by the lumican gene mutation associated with myopia

Abstract

Objective: To investigate the regulation of basic fibroblast growth factor (bFGF) and transforming growth factor (TGF)-β2 in human scleral fibroblasts (HSFs) by the adenovirus-mediated Lumican gene mutation, and to illustrate the effect of this mutation on myopia. Methods: Experimental study. The HSFs were isolated and cultured from human scleral tissues. The 3rd to 5th generation HSFs were transduced with Lumican mutant (c.596T>C) adenovirus, Lumican wild-type adenovirus, and defective adenovirus as the mutant group, wild group, and negative control group, respectively. Untransduced HSFs were defined as control group. The operation was conducted three times in each group. The expression levels of Lumican, bFGF and TGF-β2 were detected by qPCR. Statistical analysis of gene expression differences between groups was performed by fold changes. The differences were analyzed by one way ANOVA combined with LSD-t test. Results: The expressions of Lumican in the mutant group and the wild group were 103.146-fold and 398.646-fold increased compared to the control group with significant difference (t=-16.641, -21.729; P<0.05). There was no statistical difference between the negative control group and the control group (t=1.689, P>0.05). The expressions of bFGF and TGF-β2 in the mutant group were 2.812-fold and 2.346-fold increased compared to the control group with significant difference, and higher than the other groups (t=-3.921, -4.851; P<0.05). There was no significant difference among the wild group, negative control group and control group (P>0.05). Conclusions: The Lumican mutation (c.596T>C) increased the expressions of bFGF and TGF-β2 in HSFs. It indicates that the Lumican mutation (c.596T>C) may change the metabolism of extracellular matrix in the sclera by regulating bFGF and TGF-β2 to participate in scleral remodeling during the process of myopia. (Chin J Ophthalmol, 2021, 57:277-283).