title: Inhibitory effect of adenoviral vector-mediated delivery of p21WAF1/CIP1 on retinal vascular endothelial cell proliferation and tube formation in cultured Rhesus monkey cells (RF/6A).
abstract: OBJECTIVE
  To investigate the inhibitory effect(s) of adenovirus (Ad) vector-mediated delivery of p21(WAF1/CIP1) (Ad-p21) on proliferation and tube formation in Rhesus monkey choroid-retina vascular endothelial cells (RF/6A).
  
  
  METHODS
  In vitro-cultured RF/6A cells were divided into three groups: phosphate-buffered saline (PBS), Ad-p21-transfected, and negative control. Plasmid vectors were transfected via Ad-p21. The mRNA and protein expressions of p21 and cyclin-dependent kinase (CDK)2 in RF/6A cells were measured by reverse transcription-PCR (RT-PCR) and western blot analyses. Cell-cycle distributions were analyzed by flow cytometry. Matrigel was used as a matrix for endothelial cell tube formation.
  
  
  RESULTS
  Expressions of p21 mRNA and protein were greater, and expressions of CDK2 mRNA and protein lower, in the Ad-p21-transfected group than in either the PBS or negative control groups. Cell-cycle distribution analysis indicated that the proportion of G0/G1 cells was significantly higher in the Ad-p21 transfected group than in either the PBS or negative control groups (p = 0.000). There were significantly fewer endothelial cell tubes in the Ad-p21-transfected group than in either the PBS or negative control groups (p = 0.004).
  
  
  CONCLUSIONS
  Ad-p21 inhibits RF/6A cell proliferation and tube formation. The underlying mechanism to account for this may be that overexpression of p21 arrests the cell-cycle transition from the G1- to the S-phase via inhibition of CDK2 activity.
authors: Department of Ophthalmology, Tianjin Medical University General Hospital, Tianjin China.
authors: Jindong
authors: J
authors: Han
authors: Zhigang
authors: Z
authors: Yuan
authors: Hua
authors: H
authors: Yan
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002478:Cells, Cultured; D002829:Choroid / Q000166:cytology / Q000502:physiology; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics*; D042783:Endothelial Cells / Q000166:cytology* / Q000502:physiology; D005434:Flow Cytometry; D016193:G1 Phase / Q000235:genetics; D018014:Gene Transfer Techniques; D008253:Macaca mulatta; D015861:Retinal Neovascularization / Q000473:pathology / Q000628:therapy*; D012171:Retinal Vessels / Q000166:cytology* / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016196:S Phase / Q000235:genetics
chemical_list: D050759:Cyclin-Dependent Kinase Inhibitor p21