title: [Vector-mediated shRNA inhibits HIF-1alpha expression in prostate cancer cells].
abstract: OBJECTIVE
  To construct a short hairpin RNA (shRNA) vector of the hypoxia inducible factor-1alpha (HIF-1alpha), determine its inhibitory effect on the expression of the HIF-1alpha gene in PC-3M cells, and investigate its application prospects in the treatment of prostate cancer.
  
  
  METHODS
  We designed and synthesized the shRNA template sequence specific against HIF-lalpha, inserted it into the vector psilencer 2.1-U6 to generate the plasmid psilencer-HIF, transfected the recombinant plasmid into prostate cancer cell line PC-3M cells and detected the transfection efficiency by cotransfection with the pEGFP vector as well as the expression of HIF-1alpha by RT-PCR and Western blot.
  
  
  RESULTS
  The DNA sequencing analysis showed a complete consistency of the recombinant plasmid psilencer-HIF with the design. Twenty-four hours after the transfection, the rate of transfected plasmid was about (89.26 +/- 4.72)% and the vector-mediated shRNA induced RNA interference (RNAi), while 48 hours transfection reduced the HIF-1alpha mRNA and protein levels by 82.09% and 81.61% respectively (P < 0.01) in PC-3M cells.
  
  
  CONCLUSIONS
  The shRNA vector was successfully constructed, which can effectively suppress the expression of HIF-1alpha in prostate cancer cells.
authors.affiliation: Department of Urology, Beijing Chaoyang Hospital, Capital University of Medical Sciences, Beijing 100020, China. pinghaozx@yahoo.com.cn
authors.forename: Hao
authors.initials: H
authors.lastname: Ping
authors.forename: Nian-Zeng
authors.initials: NZ
authors.lastname: Xing
authors.forename: Xiao-Chun
authors.initials: XC
authors.lastname: Chen
authors.forename: Jun-Hui
authors.initials: JH
authors.lastname: Zhang
authors.lastname: Yan-Yong
authors.forename: Yong
authors.initials: Y
authors.lastname: Yang
mesh_terms: D001483:Base Sequence; D045744:Cell Line, Tumor; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics*; D008297:Male; D011471:Prostatic Neoplasms / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering; D014162:Transfection
chemical_list: D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012333:RNA, Messenger; D034741:RNA, Small Interfering