title: [Effects of anti-sense Smad4 gene on the biological characteristics of the fat-storing cell line CFSC].
abstract: OBJECTIVE
  To investigate the effects of antisense Smad(4) on the biological characteristics of the fat-storing cell line CFSC.
  
  
  METHODS
  Fat-storing cells of line CFSC from rat with liver fibrosis were cultured and transfected with 50 MOI of recombinant adenoviral vector carrying antisense Smad(4) (AdvATSmad(4)) or the control empty adenovirus (Adv0), both produced by 293 packaging cells, respectively. Two, four, and six days after the transfection the cultured cells were collected to undergo trypan blue staining and cell counting. The growth curves were drawn. The presence of antisense Smad(4) was detected by RT-PCR and Western blotting. (3)H-TdR was added into the culture media to be co-cultured for 6 hours. Then the cells were collected to examine the (3)H-TdR incorporation rate. RT-PCR and immunohistochemistry were used to examine the expression of COL1A1 and type I collagen, kinds of extracellular matrix (ECM).
  
  
  RESULTS
  Compared with the control CFSC and the Adv0-transfected CFSC cells, the cell growth curve, (3)H-TdR incorporation rate, proline incorporation rate, expression of Smad(4), and expression of extracellular matrix were markedly decreased in the AdvATSmad(4)-transfected CFSCs.
  
  
  CONCLUSIONS
  The antisense Smad(4) gene inhibits the expression of Smad(4) mRNA and protein, proline incorporation and cell growth, thus down-regulating the production of ECM. Antisense Smad(4) gene may be used as a choice of gene therapy for liver fibrosis.
authors.affiliation: Department of Hepatobiliary Surgery, People's Hospital, Peking University, Beijing 100044, China.
authors.forename: Xin-bao
authors.initials: XB
authors.lastname: Xu
authors.forename: Xi-sheng
authors.initials: XS
authors.lastname: Leng
authors.forename: Zhen-ping
authors.initials: ZP
authors.lastname: He
authors.forename: Zhi-qing
authors.initials: ZQ
authors.lastname: Liang
authors.forename: Kai
authors.initials: K
authors.lastname: Lin
authors.forename: Xin
authors.initials: X
authors.lastname: Yu
authors.forename: Yu-hua
authors.initials: YH
authors.lastname: Wei
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D002455:Cell Division; D002460:Cell Line; D024042:Collagen Type I / Q000378:metabolism; D016373:DNA, Antisense / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D008099:Liver / Q000378:metabolism / Q000473:pathology; D011392:Proline / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051901:Smad4 Protein; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D024042:Collagen Type I; D016373:DNA, Antisense; D004268:DNA-Binding Proteins; D051901:Smad4 Protein; C490811:Smad4 protein, rat; D015534:Trans-Activators; D011392:Proline