title: Effects of shRNA-silenced livin and survivin on lung cancer cell proliferation and apoptosis.
abstract: OBJECTIVE
  To evaluate the effects of short hairpin RNA (shRNA)-mediated silencing of livin and survivin on the proliferation and apoptosis of A549 lung cancer cells.
  
  
  METHODS
  We designed and constructed the eukaryotic expression vectors pSilencer-livin and pSilencer-survivin which contain the Livin and Survivin genes, respectively, and transfected them into liposome-combined lung cancer A549 cells. The cells were then divided into the blank control, plasmid control, Livin, Survivin, and co-transfected groups. Real-time quantitative PCR (qRT-PCR) and Western blot assay were used to determine the mRNA and protein expression levels of Livin and Survivin. The MTT assay was used to evaluate the changes in cell proliferation. The TUNEL assay was used to evaluate the apoptotic rate.
  
  
  RESULTS
  The shRNA eukaryotic expression vectors of Livin and Survivin were successfully constructed. The mRNA and protein expression of Livin and Survivin were significantly lower in the co-transfected group than in the control groups (p<0.05) . At 48, 60, and 72 hrs after transfection, the cell growth inhibition rate was significantly higher in the co-transfected group than in the single- transfected group (p<0.05) . At 48 hrs after transfection, the apoptotic rate significantly increased (p<0.05) .
  
  
  CONCLUSIONS
  Co-silencing of Livin and Survivin can effectively inhibit the cell proliferation and apoptosis of lung cancer cells.
authors.affiliation: Department of Respiratory Medicine, the First Affiliated Hospital of Xinxiang Medical University, Weihui, P.R., China.
authors.forename: Xin
authors.initials: X
authors.lastname: Zhao
authors.forename: Yan
authors.initials: Y
authors.lastname: Yuan
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Zhang
authors.forename: Xianjun
authors.initials: X
authors.lastname: Feng
authors.forename: Jian
authors.initials: J
authors.lastname: Zhang
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Yuan
authors.forename: Jinlan
authors.initials: J
authors.lastname: Li
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000502:physiology*; D017209:Apoptosis; D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D020868:Gene Silencing; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics / Q000502:physiology*; D008175:Lung Neoplasms / Q000473:pathology*; D009363:Neoplasm Proteins / Q000235:genetics / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D000077022:Survivin
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C107489:BIRC5 protein, human; C419866:BIRC7 protein, human; D051034:Inhibitor of Apoptosis Proteins; D009363:Neoplasm Proteins; D034741:RNA, Small Interfering; D000077022:Survivin