title: Biologically inhibitory effects of VEGF siRNA on endometrial carcinoma cells.
abstract: OBJECTIVE
  To study the biological effects of small inhibitory RNA (siRNA) on endometrial carcinoma cells through disrupting the mRNA of Vascular endothelial growth factor (VEGF), and its inhibitory functions on tumor cells' proliferation.
  
  
  METHODS
  The sequence-specific siRNA of VEGF165 and the mock siRNA were designed, they were transfected into the endometrial carcinoma cell line, Ishikawa cells. Total cell RNAs were extracted from the transfected cells. The mRNA levels of VEGF were then analyzed utilizing real-time fluorescence quantitative RT-PCR (FQ-PCR) post-transfection at 12, 24, 48, 72, and 168 h, respectively. Simultaneously its protein levels were also determined by western blot. Cell proliferations were monitored by MTT assay and cytopathological effects (CPE) were determined by Multispect Imaging System.
  
  
  RESULTS
  The mRNA and protein levels of VEGF declined post-transfection at 12, 24, 48, 72 h, respectively, with a maximum decrease post-transfection at 48 h. The proliferation of Ishikawa cells was significantly inhibited in a similar manner during this time window. All the data were repeated at least three independent times and similar results were achieved.
  
  
  CONCLUSIONS
  The small siRNA of VEGF165 can effectively down-regulate its target mRNA and protein levels; consequently proliferation of Ishikawa cells was inhibited. The data strongly imply that VEGF may be involved in the genesis of endometrial carcinoma, which provides a new pathway to treat human endometrial caners.
authors: Department of Obstetrics and Gynecology, Affiliated Hospital of Qingdao University School of Medicine, Qingdao 266003, People's Republic of China. shuping.zhao@yahoo.com
authors: Shuping
authors: S
authors: Zhao
authors: Dehua
authors: D
authors: Ma
authors: Hongying
authors: H
authors: Dai
authors: Fang
authors: F
authors: Yuan
authors: Zenyan
authors: Z
authors: Wang
mesh_terms: D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015536:Down-Regulation; D016889:Endometrial Neoplasms / Q000235:genetics* / Q000378:metabolism; D005260:Female; D020868:Gene Silencing / Q000502:physiology; D006801:Humans; D015345:Oligonucleotide Probes / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism
chemical_list: D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A