title: [Effects of siRNAs silencing transforming growth factor-beta1 on fibronectin in SD rat masangial cells].
abstract: OBJECTIVE
  To investigate fibronectin synthesis in SD rat mesangial cells after transforming growth factor-beta1 (TGF-beta1) is silenced by the short interfering RNA (siRNA) expressed by reconstructed pGEFP-C1 vectors.
  
  
  METHODS
  Depending upon the 538th - 556th (A) and 895th - 913th (B) nucleotides of rat TGF-beta1 gene, a nucleotide (A or B) was constructed into a small hairpin nucleotide which was separately (A or B) or together (A plus B) inserted into a pGEFP-C1 vector with three reconstructed pGEFP-C1 vectors separately expressing the siRNAs for A or/and B. TGF-beta1 and fibronectin were dynamically investigated for their interrelationship by ELISA in the supernatant and RT-PCR in their extracted total RNA.
  
  
  RESULTS
  The siRNA hairpin-like molecules were constructed according to the 538th - 556th nucleotides of rat TGF-beta1 gene were able to markedly silence the expression of TGF-beta1 mRNA (P < 0.01) and protein (P < 0.01) at 48 h. Lipfectamin 2000 transfection stimulated the peak secretion of fibronectin at 24 h in the control and the experimental group whose TGF-beta1 was not silenced, but the silence of TGF-beta1 in both experimental groups delayed the top values of fibronectin to 48 h (P < 0.01).
  
  
  CONCLUSIONS
  The silence of TGF-beta1 by siRNA decreased the fibronectin expression, but the latter was possibly not completely TGF-dependent.
authors.affiliation: Clinical Center of Pediatric Nephrology, The Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Hua-xiong
authors.initials: HX
authors.lastname: Mao
authors.forename: Zhu-wen
authors.initials: ZW
authors.lastname: Yi
authors.forename: Xiao-chuan
authors.initials: XC
authors.lastname: Wu
authors.forename: Xi-qiang
authors.initials: XQ
authors.lastname: Dang
authors.forename: Xiao-jie
authors.initials: XJ
authors.lastname: He
authors.forename: Yan
authors.initials: Y
authors.lastname: Cao
authors.forename: Shuang-hong
authors.initials: SH
authors.lastname: Mo
mesh_terms: D000818:Animals; D002477:Cells; D002478:Cells, Cultured; D005353:Fibronectins / Q000378:metabolism*; D050527:Mesangial Cells / Q000187:drug effects* / Q000378:metabolism; D034622:RNA Interference / Q000187:drug effects* / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D053773:Transforming Growth Factor beta1 / Q000737:chemistry / Q000235:genetics*
chemical_list: D005353:Fibronectins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D053773:Transforming Growth Factor beta1