title: [Small interfering RNA inhibits the expression of connexin43 in the human corpus cavernosum penis smooth muscle cells].
abstract: OBJECTIVE
  To inhibit the expression of connexin43 (Cx43) in the human corpus cavernosum penis smooth muscle cells by small interfering RNA (siRNA) and detect the gap junction intercellular communication (GJIC), and to investigate the application of siRNA technology in the gap junction of corpus cavernosum penis smooth muscle cells and its role in the penile erection process.
  
  
  METHODS
  With the help of the software of Ambion Corporation, the specific recombinant plasmids with siRNA targeting human Cx43 gene were constructed. The recombinant plasmids having been stably transferred into human corpus cavernosum penis smooth muscle cells for 48 hours, semi-quantitive reverse transcription polymerase chain reaction (RT-PCR) and Western blotting techniques were used to examine the inhibitory effects of siRNA on the expressions of the Cx43 gene and protein, in comparison with the siRNA negative control and the blank control group, respectively. The GJIC was detected by scrape-loading and fluorescence dye transfer experiments through the fluorescence microscope.
  
  
  RESULTS
  The results of enzyme digestion analysis and DNA sequencing showed that the recombinant plasmid pSilencer 1.0-U6-siRNA-Cx43 was successfully constructed. The relative levels of Cx43 mRNA and protein expression in the smooth muscle cells were (0.45 +/- 0.08)% and (0.56 +/- 0.06)% after successful transfer of the recombinant plasmid. However, the expression levels of mRNA and protein were (0.72 +/- 0.04)% and (0.80 +/- 0.08)% in the negative siRNA transfer group, and (0.74 +/- 0.09)% and (0.77 +/- 0.11)% in the blank control, respectively, with a significant difference (P < 0.05). The GJIC also decreased significantly.
  
  
  CONCLUSIONS
  siRNA can significantly inhibit the expression of Cx43 and block the GJIC in the human corpus cavernosum penis smooth muscle cells. siRNA technology plays an important role in penile erection and flaccidity.
authors.affiliation: Department of Urology, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, Anhui 230001, China. zgcao2002@126.com
authors.forename: Zheng-guo
authors.initials: ZG
authors.lastname: Cao
authors.forename: Yu-ping
authors.initials: YP
authors.lastname: Zhu
authors.forename: You-wen
authors.initials: YW
authors.lastname: Sun
authors.forename: Xiao-cheng
authors.initials: XC
authors.lastname: Dong
authors.forename: Lin
authors.initials: L
authors.lastname: Qi
authors.forename: Jun
authors.initials: J
authors.lastname: Xiao
authors.forename: Hao
authors.initials: H
authors.lastname: Chen
authors.forename: Wei-hua
authors.initials: WH
authors.lastname: Ren
authors.forename: Lian
authors.initials: L
authors.lastname: Zou
mesh_terms: D015152:Blotting, Northern; D002478:Cells, Cultured; D018031:Connexin 43 / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D007365:Intercellular Junctions; D008297:Male; D032389:Myocytes, Smooth Muscle / Q000502:physiology*; D010413:Penis / Q000166:cytology / Q000378:metabolism*; D034741:RNA, Small Interfering; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
chemical_list: D018031:Connexin 43; D034741:RNA, Small Interfering