Effect of gene silencing of Golgi a-mannosidase . on the invasion and migration of human gastric carcinoma BGC-823 cells
10.3781/j.issn.1000-7431.2012.06.002
- Author:
Ya-Ying YANG
1
Author Information
1. Department of Pathology
- Publication Type:Journal Article
- Keywords:
Cell migration assays;
Gene silencing;
Mannosidases;
Neoplasm invasiveness;
Stomach neoplasms
- From:
Tumor
2012;32(6):402-407
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effect of gene silencing of Golgi a-mannosidase II(GM II) by RNA interference on the invasion and migration of human gastric carcinoma BGC-823 cells. Methods: Four targeted locations were designed and four plasmid vectors expressing short hairpin RNA (shRNA) for gene silencing of GM II were constructed, and the negative control plasmid vector was also constructed. The plasmids were transfected into human gastric carcinoma BGC-823 cells by Lipofectamine 2000. The expression levels of GMIImRNA and protein were detected by RT-PCR and Western blotting, respectively. The best plasmid for gene silencing of GMIIwas selected. The invasive and migration capabilities of BGC-823 cells induced by gene silencing of GMIIwere detected by Transwell invasion assay and scratch-wound assay, respectively. Results: Plasmid vector pGPU6/GFP/Neo-1303 was the best one for the gene silencing of GM .. The number of cells invading through the matrigel in the plasmid vector pGPU6/GFP/Neo-1303-transfected group was much lower than those in the blank control group and the pGPU6/GFP/Neo-shNC-transfected group (P < 0.05). After the cells were cultured without serum for 24 h, the invasive and migration capabilities of the cells transfected with pGPU6/GFP/Neo-1303 were significantly weakened than those of the blank control group and the pGPU6/GFP/Neo-shNC- transfected group (P < 0.05). Conclusion: The expressions of GMIImRNA and protein in human gastric carcinoma BGC-823 cells can be silenced by RNA interference, which results in the inhibition of invasive and migration capabilities of BGC-823 cells. These results suggest that GMIImight be a novel target for the treatment of human gastric carcinoma. © 2012 by Tumor.
