title: Silencing of twist expression by RNA interference suppresses epithelial-mesenchymal transition, invasion, and metastasis of ovarian cancer.
abstract: OBJECTIVE
  This study aimed to explore the role of the Twist gene in the epithelial-mesenchymal transition of ovarian cancer.
  
  
  METHODS
  An RNA interference plasmid expressing a small interfering RNA (siRNA)-targeting Twist (Twist siRNA vector) was designed, constructed, and transfected into the human ovarian cancer cell line A2780. Transfection efficiency was assessed under a fluorescence microscope. Changes in the expression of Twist mRNA in A2780 after transfection with the pGenesil Twist shRNA plasmid were analyzed through RT-PCR. MTT assays and adhesion experiments were applied to determine changes in proliferation and adhesion ability of A2870 after transfection with the Twist shRNA plasmid. Changes in the expression of the E-cadherin and N-cadherin proteins in A2780 after transfection with the Twist shRNA plasmid were analyzed using Western blotting.
  
  
  RESULTS
  The restructuring plasmid pGenesil-Twist shRNA was constructed successfully. After 48 h of culture, 80% of the cells expressed high-intensity GFP fluorescence and stability. The expression of Twist decreased significantly after the transfection of the Twist shRNA plasmid (P<0.05). Proliferation of the transfected Twist shRNA cells showed no difference with that of the A2780-nontransfection or A2780-si-control groups (P>0.05) but the adhesion ability of A2780 decreased dramatically (P<0.05). Expression of the E-cadherin protein increased, whereas that of the N-cadherin protein decreased compared with that in the A2780-nontransfection or A2780- si-control groups (P<0.05).
  
  
  CONCLUSIONS
  Twist is essential for epithelial-mesenchymal transition, invasion, and metastasis of ovarian cancer.
authors.affiliation: Department of Gynecology, Yantai Yuhuangding Hospital, Yantai, China.
authors.forename: Wen-Shuang
authors.initials: WS
authors.lastname: Wang
authors.forename: Xing-Sheng
authors.initials: XS
authors.lastname: Yang
authors.forename: Min
authors.initials: M
authors.lastname: Xia
authors.forename: Hai-Yang
authors.initials: HY
authors.lastname: Jiang
authors.forename: Jian-Qing
authors.initials: JQ
authors.lastname: Hou
mesh_terms: D015820:Cadherins / Q000378:metabolism; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering; D014162:Transfection; D051879:Twist-Related Protein 1 / Q000235:genetics* / Q000378:metabolism
chemical_list: D015820:Cadherins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051879:Twist-Related Protein 1; D049452:Green Fluorescent Proteins