title: Effect of siRNA on Wisp-1 gene expression, proliferation, migration and adhesion of mouse hepatocellular carcinoma cells.
abstract: OBJECTIVE
  To study the inhibition effect of siRNA on the expression of Wisp-1 gene in Hca-F of mouse hepatocellular carcinoma cells strain and also its effect on the proliferation, migration and adhesion of hepatocellular carcinoma cells.
  
  
  METHODS
  Three expression vectors of siRNA were constructed. Lipo2000 was employed to transfect Hca-F cells and Western blot was used to detect the inhibition effect of siRNA on the expression of Wisp-1 gene. Afterward, CCK8 was adopted to detect the effect of Wisp-1 siRNA on the proliferation of Hca-F cells; Annexin V-FITC/PI double staining flow cytometry was used to detect the effect of Wisp-1 siRNA on the apoptosis of Hca-F cells; Transwell was used to detect the effect of Wisp-1 siRNA on the migration of Hca-F cells. The in vitro cell adhesion kit was used to detect of Wisp-1 siRNA on the change in the components of extracellular matrix to which Hca-F cells adhered. Western blot was used to detect the activation of protein kinase B (AKT)/glycogen synthase kinase-3β pathway and the expression of downstream target protein p53 and matrix metalloproteinases-2.
  
  
  RESULTS
  The siRNA showed interference effect on the expression of Wisp-1 gene. Compared with the control group, after being transfected to cells, Wisp-1 siRNA could significantly inhibit the proliferation, migration and adhesion of Hca-F cells and also promote the cell apoptosis, which was related to the down-regulated phosphorylation of AKT and glycogen synthase kinase-3β and the expression of p53 and matrix metalloproteinases-2 (P < 0.05).
  
  
  CONCLUSIONS
  The inhibition of Wisp-1 expression can reduce the proliferation, migration and adhesion of mouse hepatocellular carcinoma cells, which is related to the AKT/glycogen synthase kinase-3β pathway. Wisp-1 gene may be the potential target to cure the hepatocellular carcinoma.
authors.affiliation: Department of Gastroenterology, Affiliated Shandong Provincial Hospital of Shandong University, Ji'nan 250012, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Ge
authors.affiliation: Department of Gastroenterology, Affiliated Shandong Provincial Hospital of Shandong University, Ji'nan 250012, China.
authors.forename: Xiao-Hua
authors.initials: XH
authors.lastname: Zhang
authors.affiliation: School of Nursing, Binzhou Vocational College, Binzhou 256603, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.affiliation: Department of Gastroenterology, Zhangqiu People's Hospital, Zhangqiu 250200, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Gastroenterology, Zhangqiu People's Hospital, Zhangqiu 250200, China.
authors.forename: Qing-Yan
authors.initials: QY
authors.lastname: Li
authors.affiliation: Department of Gastroenterology, General Hospital of Ningxia Medical University, Yinchuan 750004, China. Electronic address: nyfytaowei@163.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Tao
authors.affiliation: Department of Medical Oncology, Shandong Cancer Hospital and Institute, Ji'nan 250117, China. Electronic address: rghhappiness@126.com.
authors.forename: Guo-Hua
authors.initials: GH
authors.lastname: Ren
keywords: AKT; Hepatocellular carcinoma cells; RNA interference; Wisp-1