title: Small interfering RNA transfection against serum response factor mediates growth inhibition of benign prostatic hyperplasia fibroblasts.
abstract: To treat urethral strictures of the lower urinary tract, urethrotomy is the method of choice. But this minimally invasive method suffers from poor outcome rates and leads often to restenosis of the urinary tract because of hyper-proliferating fibroblasts. Our aim is to minimize the proliferation of excessive tissue due to a new minimal invasive therapeutic approach. As an appropriate model, we isolated fibroblasts from different benign prostatic hyperplasia patients and transfected them with small interfering RNA (siRNA) against the transcription factor serum response factor (SRF), a key factor for cell cycle regulation and apoptosis. The resulting knockdown of SRF was examined on the messenger RNA level by quantitative real-time polymerase chain reaction and on the protein level by western blot. The correlation of SRF silencing and impact on cell proliferation was examined by xCELLigence, 5-bromo-2'-deoxiuridine proliferation assay, total cell counts, and senescence assay. The transfection of primary prostatic fibroblasts with SRFsiRNA revealed specific and significant knockdown of SRF, leading to significant inhibition of proliferation after the second transfection, which was revealed by proliferation assay and total cell number. The results of this study indicate a substantial role of SRF in prostatic fibroblasts and we suggest that SRF silencing might be used for the treatment of urethral strictures to achieve a durably patent urethra.
authors: Department of Thoracic, Cardiac, and Vascular Surgery, Tuebingen University Hospital, Tuebingen, Germany.
authors: Andrea
authors: A
authors: Nolte
authors: Stefan
authors: S
authors: Aufderklamm
authors: Katrin
authors: K
authors: Scheu
authors: Tobias
authors: T
authors: Walker
authors: Olivia
authors: O
authors: König
authors: Miriam
authors: M
authors: Böttcher
authors: Jan
authors: J
authors: Niederlaender
authors: Christian
authors: C
authors: Schwentner
authors: Christian
authors: C
authors: Schlensak
authors: Arnulf
authors: A
authors: Stenzl
authors: Hans Peter
authors: HP
authors: Wendel
mesh_terms: D049109:Cell Proliferation; D002478:Cells, Cultured; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008297:Male; D011470:Prostatic Hyperplasia / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics* / Q000627:therapeutic use*; D026362:Serum Response Factor / Q000037:antagonists & inhibitors* / Q000235:genetics; D014162:Transfection
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; C476858:SRF protein, human; D026362:Serum Response Factor