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
Hauptverfasser: Jing, Jiaona, Li, Ping, Li, Tiejun, Sun, Yuncheng, Guan, Huaijin
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Li, Ping
Li, Tiejun
Sun, Yuncheng
Guan, Huaijin
description 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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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).</description><identifier>ISSN: 2090-004X</identifier><identifier>DOI: 10.1155/2013/354798</identifier><language>eng</language><publisher>John Wiley &amp; Sons, Inc</publisher><subject>Analysis ; Bone morphogenetic proteins ; Care and treatment ; Diagnosis ; Genetic engineering ; Glaucoma ; Health aspects ; RNA ; Transforming growth factors</subject><ispartof>Journal of ophthalmology, 2013-01</ispartof><rights>COPYRIGHT 2013 John Wiley &amp; Sons, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,860,27903,27904</link.rule.ids></links><search><creatorcontrib>Jing, Jiaona</creatorcontrib><creatorcontrib>Li, Ping</creatorcontrib><creatorcontrib>Li, Tiejun</creatorcontrib><creatorcontrib>Sun, Yuncheng</creatorcontrib><creatorcontrib>Guan, Huaijin</creatorcontrib><title>RNA interference targeting connective tissue growth factor inhibits the transforming growth factor-[β.sub.2] induced proliferation in human tenon capsule fibroblasts</title><title>Journal of ophthalmology</title><description>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. 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subjects Analysis
Bone morphogenetic proteins
Care and treatment
Diagnosis
Genetic engineering
Glaucoma
Health aspects
RNA
Transforming growth factors
title RNA interference targeting connective tissue growth factor inhibits the transforming growth factor-[β.sub.2] induced proliferation in human tenon capsule fibroblasts
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