TPX2 gene-targeted short hairpin RNA induces the apoptosis of human lung adenocarcinoma A549 cells and its related mechanism
10.3781/j.issn.1000-7431.2011.12.001
- Author:
Ying LIU
1
Author Information
1. Department of Pathology
- Publication Type:Journal Article
- Keywords:
Apoptosis;
Gene, TPX 2;
Lung neoplasms;
RNA interference
- From:
Tumor
2011;31(12):1055-1060
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effect of short hairpin RNA (shRNA) targeting Xenopus kinesin-like protein 2 (TPX 2) gene on the apoptosis of human lung adenocarcinoma A549 cells and to explore its possible mechanism. Methods: The combination vector of shRNA targeting TPX2 gene was established and then it was transfected into A549 cells. The expression levels of TPX2, Aurora-A, p53 and Bcl-2 mRNAs in A549 cells after transfection with combination vector were detected by RT-PCR, and the expression level of TPX2 protein was determined by Western-blotting. The cell cycle distribution and the apoptosis were analyzed by flow cytometry (FCM). Results: The combination vector of pMagic4.1-shRNA-TPX2 was successfully established. The expression levels of TPX2, Aurora-A and Bcl-2 mRNAs were down-regulated whereas the expression level of p53 mRNA was up-regulated in A549 cells after transfection with pMagic4.1-shRNA-TPX2, and the expression level of TPX2 protein was down-regulated. The apoptosis rate of A549 cells was increased after transfection with pMagic4.1-shRNA-TPX2, and the cell cycle was arrested at S phase. These changes in the pMagic4.1-shRNA-TPX2- transfected group were significantly different from those in the blank control group (without transfection with combination vector) and the negative control group (transfection with pMagic4.1-shRNA-NC) (P<0.05). Conclusion: The shRNA targeting TPX 2 gene can induce the apoptosis of A549 cells, and this effect may be related to up-regulation of p53 expression and down-regulation of Bcl-2 expression. Copyright© 2011 by TUMOR.
