title: Up-regulation of VEZT by small activating RNA inhibits the proliferation, invasion and migration of gastric cancer cells.
abstract: OBJECTIVE
  To identify an effective saRNA sequence that can specifically up-regulate VEZT expression and to determine the influence of saRNA had on gastric cancer cell growth, proliferation, invasion and migration.
  
  
  METHODS
  Three various saRNAs, that target the VEZT gene promoter at different locations relative to the transcription start site were synthesized. A dsControl saRNA was synthesized as a negative control, and a specific shRNA was synthesized to knockdown VEZT and eliminate any off-target effects of the saRNA. Both SGC-7901 and M-28 cells were either transfected with the different saRNAs, or treated with Lipofectamine2000 alone. To determine the most effective saRNA, real-time PCR and Western blot were used to determine the VEZT mRNA and protein content, respectively, of each treatment group. After selection, both cell lines were treated with the chosen saRNA, dsControl or Lipofectamine2000. The saRNA treated cells were divided into two groups: the first group was used immediately in the experiments, and the second group was transfected with shRNA by using RNAi-Mate. The proliferation of cells transfected with saRNA, or saRNA and shRNA, as well as the other control cells, was detected by CCK-8. The invasive and migratory abilities were determined using the transwell chamber assay.
  
  
  RESULTS
  We identified the most effective saRNA via real-time PCR and Western blot. The selected saRNA inhibited the growth, invasion and migration of GC cells by specially reactivating VEZT. The real-time PCR and Western blot results showed that treatment with saRNA caused a significant up-regulation of VEZT, and an obvious decrease in the proliferative, invasive and migratory abilities; compared with the control groups (P < 0.01); furthermore, there were no significant differences among the control groups (P > 0.05). This phenomenon provides a theoretical basis for saRNA design and gene therapy for gastric cancer.
authors.affiliation: Department of Gastrointestinal Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250000, Shandong, China.
authors.forename: Detian
authors.initials: D
authors.lastname: Xie
authors.affiliation: Department of Gastrointestinal Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250000, Shandong, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Shang
authors.affiliation: Department of Gastrointestinal Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250000, Shandong, China.
authors.forename: Lipan
authors.initials: L
authors.lastname: Peng
authors.affiliation: Department of Gastrointestinal Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250000, Shandong, China. Electronic address: doclileping@163.com.
authors.forename: Leping
authors.initials: L
authors.lastname: Li
mesh_terms: D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D009361:Neoplasm Invasiveness / Q000473:pathology; D012313:RNA / Q000008:administration & dosage / Q000235:genetics*; D034741:RNA, Small Interfering; D013270:Stomach / Q000166:cytology / Q000473:pathology*; D013274:Stomach Neoplasms / Q000235:genetics* / Q000473:pathology* / Q000628:therapy; D015854:Up-Regulation
chemical_list: D002352:Carrier Proteins; D008565:Membrane Proteins; D034741:RNA, Small Interfering; C494547:VEZT protein, human; D012313:RNA
keywords: Gastric cancer; Invasion; Migration; VEZT; saRNA