RNA Interference targeting connective tissue growth factor inhibits the transforming growth factor- β 2 induced proliferation in human Tenon capsule fibroblasts

Purpose. this study was to determine the effect of ctgf-small interfering rna (sirna) on tgf-β2-induced proliferation in human tenon capsule fibroblasts (htfs). methods. htfs were transfected with four of ctgf-sirnas separately for screening of gene silencing efficacy that was determined by transcript level measured by quantitative real-time pcr (qrt-pcr). recombinant tgf-β2 was added into the culture to stimulate the proliferation of htfs. the gene silencing efficacy of the sirnas was evaluated by qrt-pcr and immunofluorescence of ctgf transcript and protein levels. the viability of htfs was determined by cell counting kit-8 (cck-8). fcm was used to assess cell cycle after ctgf-sirna transfection. results. the expression of ctgf and proliferation of htfs were increased significantly by tgf-β2 stimulation. the transfection of ctgf-sirna abolished the upregulation of ctgf and cell proliferation induced by tgf-β2. the analysis of cell cycle indicated that ctgf-sirna treatment stimulated cells from s phase to g0/g1 phase in comparison with the inverse physiologic function of tgf-β2. conclusion. ctgf targeting sirna could effectively suppress the expression of ctgf and attenuate the proliferation of htfs. the sirna approach may provide a therapeutic option for eliminating filtration bleb scarring after glaucoma filtration surgery (gfs). © 2013 jiaona jing et al.