title: [Effect of antisense thrombin receptor and p21 double gene co-expression system on the proliferation and apoptosis in human aortic smooth muscle cells].
abstract: OBJECTIVE
  To focus on the study of the effect on proliferation and apoptosis of human aortic smooth muscle cells (ASMC) by adeno-associated virus (AAV) vector carrying antisense thrombin receptor (ATR) and p21 double gene co-expression system.
  
  
  METHODS
  Cultured human AMSC was infected with recombinant AAV containing ATR, p21 single gene and AP double gene respectively. The integration and expression of genes were confirmed by semi-quantitative RT-PCR. The anti-proliferation effect was determined by MTT assay. Cell cycle and apoptotic cell counts were measured through Flow Cytometry. The rate of apoptotic cells was examined with acridine orange/ethidium bromide(AO/EB) stain.
  
  
  RESULTS
  RT-PCR indicated that the exogenous genes had been integrated into ASMC. The rates of cell survival were decreased by 16.67%, 21.60%, and 29.4% and the cell counts of G0/G1 phase were (61.8 +/- 2.9)%, (82.5 +/- 4.0)%, (80.4 +/- 6.1)% in ATR, p21 and AP group respectively after rAAV infected 4 days. The level and area of apoptotic peak were greater in AP double gene than ATR and p21 single gene. Cell stain indicated that apoptotic cells were (7.2 +/- 3.3)%, (10.7 +/- 5.6)%, and (18.3 +/- 2.7)% in each transgene group compared with (1.5 +/- 0.8)% in control group.
  
  
  CONCLUSIONS
  AP double gene co-expression system has powerful effect for inhibiting proliferation and inducing apoptosis ASMC than ATR and p21 single gene and that is a superior way for gene therapy to restenosis.
authors.affiliation: Molecular Medical Center, Cardiovascular Institute, Fu Wai Hospital, CAMS, PUMC, Beijing 100037, China. xmmengcn@yahoo.com
authors.forename: Xian-min
authors.initials: XM
authors.lastname: Meng
authors.forename: Li-guo
authors.initials: LG
authors.lastname: Mi
authors.forename: Xiu-wen
authors.initials: XW
authors.lastname: Zhao
authors.forename: Hui-qing
authors.initials: HQ
authors.lastname: Cao
authors.forename: Yun-lin
authors.initials: YL
authors.lastname: Gao
authors.forename: Jin-feng
authors.initials: JF
authors.lastname: Ding
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics; D016375:Antisense Elements (Genetics); D001011:Aorta / Q000166:cytology; D017209:Apoptosis; D002455:Cell Division; D002478:Cells, Cultured; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000096:biosynthesis / Q000235:genetics*; D005333:Fetus; D005822:Genetic Vectors; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D018179:Receptors, Thrombin / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D016375:Antisense Elements (Genetics); C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D018179:Receptors, Thrombin