RNA interference targeting connective tissue growth factor inhibits the transforming growth factor-[β.sub.2] induced proliferation in human tenon capsule fibroblasts
Purpose: This study was to determine the effect of CTGF-small interfering RNA (siRNA) on TGF-[β.sub.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 t...
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Veröffentlicht in: | Journal of ophthalmology 2013-01 |
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Sprache: | eng |
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Zusammenfassung: | Purpose: This study was to determine the effect of CTGF-small interfering RNA (siRNA) on TGF-[β.sub.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 TGFwas 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.sub.2] stimulation. The transfection of CTGF-siRNA abolished the upregulation of CTGF and cell proliferation induced by TGF-[β.sub.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.sub.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). |
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ISSN: | 2090-004X |
DOI: | 10.1155/2013/354798 |