Targeted gene silencing of homo sapiens eukaryotic translation elongation factor 1 alpha 2 inhibits migration and invasion of human pancreatic cancer cell line BxPC-3 in vitro
10.3781/j.issn.1000-7431.2012.08.001
- Author:
Chao XU
1
Author Information
1. Department of Gastroenterology
- Publication Type:Journal Article
- Keywords:
Cell migration assays;
Homo sapiens eukaryotic translation elongation factor 1 alpha 2;
Neoplasm invasiveness;
Pancreatic neoplasms;
Peptide elongation factors;
RNA interference
- From:
Tumor
2012;32(8):567-571
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effect of targeted gene silencing of homo sapiens EEF1A2 (eukaryotic translation elongation factor 1 alpha 2) on migration and invasion of human pancreatic cancer cell line BxPC-3 in vitro , and to explore its possible molecular mechanism. Methods: The mRNA and protein expression levels of EEF1A2 in four strains of human pancreatic cancer cells were detected by RT-PCR and Western blotting, respectively. EEF1A2-siRNA (small interference RNA) and negative control siRNA were transfected into BxPC-3 cells, respectively. Then the mRNA and protein expressions of EEF1A2 were determined by RT-PCR and Western blotting, respectively. The abilities of migration and invasion of BxPC-3 cells were determined by wound healing assay and Transwell invasion assay, respectively. The changes of expressions of p-Akt and total Akt proteins were detected by Western blotting. Results: Of the four strains of human pancreatic cancer cells, SW1990 cells displayed a low level of EEF1A2, and the remaining three strains of human pancreatic cancer cells including BxPC-3, Patu8988 and Panc-1 all displayed high expressions of EEF1A2. At 48 h post-transfection, the expression levels of EEF1A2 mRNA and protein in BxPC-3 cells transfected with EEF1A2-siRNA were significantly decreased as compared with those transfected with negative control siRNA or without any transfection (P < 0.01). The abilities of migration and invasion of the BxPC-3 cells transfected with EEF1A2-siRNA were obviously inhibited as compared with those transfected with negative control siRNA or without any transfection (P < 0.05). The phosphorylation of Akt protein was also significantly decreased in BxPC-3 cells transfected with EEF1A2-siRNA (P < 0.05), while the total Akt protein expression level was similar among different transfection groups (P > 0.05). Conclusion: Targeted silencing of EEF 1A 2 by siRNA can obviously inhibit the migration and invasion abilities of human pancreatic cancer BxPC-3 cells in vitro . This effect may be associated with Akt signaling pathway regulated by EEF1A2. Copyright © 2012 by TUMOR.
