Effect of siRNA on Wisp-1 gene expression, proliferation, migration and adhesion of mouse hepatocellular carcinoma cells
10.1016/j.apjtm.2015.09.008
- Author:
Jian GE
1
;
Xiao-Hua ZHANG
1
;
Fang WANG
2
;
Yu WANG
3
;
Qing-Yan LI
3
;
Guo-Hua REN
4
;
Wei TAO
5
Author Information
1. Department of Gastroenterology, Affiliated Shandong Provincial Hospital of Shandong University
2. School of Nursing, Binzhou Vocational College
3. Department of Gastroenterology, Zhangqiu People's Hospital
4. Department of Medical Oncology, Shandong Cancer Hospital and Institute
5. Department of Gastroenterology, General Hospital of Ningxia Medical University
- Publication Type:Journal Article
- Keywords:
AKT;
Hepatocellular carcinoma cells;
RNA interference;
Wisp-1
- From:
Asian Pacific Journal of Tropical Medicine
2015;8(10):821-828
- CountryChina
- Language:Chinese
-
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.
