title: RNA Interference Targeting Connective Tissue Growth Factor Inhibits the Transforming Growth Factor- β 2 Induced Proliferation in Human Tenon Capsule Fibroblasts.
abstract: Purpose. This study was to determine the effect of CTGF-small interfering RNA (siRNA) on TGF- β 2-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- β 2 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- β 2 stimulation. The transfection of CTGF-siRNA abolished the upregulation of CTGF and cell proliferation induced by TGF- β 2. 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- β 2. 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).
authors.affiliation: Eye Institute, Affiliated Hospital of Nantong University, 20 Xisi Road, Nantong, Jiangsu Province 226001, China ; Nanjing Governmental Hospital, 116 Chengxian Street, Nanjing, Jiangsu Province 210018, China.
authors.forename: Jiaona
authors.initials: J
authors.lastname: Jing
authors.forename: Ping
authors.initials: P
authors.lastname: Li
authors.forename: Tiejun
authors.initials: T
authors.lastname: Li
authors.forename: Yuncheng
authors.initials: Y
authors.lastname: Sun
authors.forename: Huaijin
authors.initials: H
authors.lastname: Guan