1. [Expression and localization of adenovirus-mediated transcriptional factor Runx3 in malignant glioblastoma cells].
- Author
-
Bai WD, Zhang R, Meng YL, Cao YX, Wang T, Zhao J, and Yang AG
- Subjects
- Animals, Cell Line, Tumor, Cell Nucleus metabolism, Cloning, Molecular, Core Binding Factor Alpha 3 Subunit biosynthesis, Core Binding Factor Alpha 3 Subunit isolation & purification, Gene Expression, Humans, Mice, Plasmids genetics, Protein Transport, Recombinant Fusion Proteins biosynthesis, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins isolation & purification, Recombinant Fusion Proteins metabolism, Adenoviridae genetics, Core Binding Factor Alpha 3 Subunit genetics, Core Binding Factor Alpha 3 Subunit metabolism, Glioblastoma pathology
- Abstract
Aim: To construct the replicative deficient adenovirus Ad-Runx3 expressing Runx3, and to express it in U251 malignant glioblastoma cells., Methods: The runx3 gene with a flag tag was amplified by PCR using pCMV5-AML2 as a template, and was confirmed by DNA sequencing. The adenovirus shuttle vector pShuttle-CMV-Runx3 was constructed by introducing runx3 DNA fragment into the sites of Kpn I and Xho I of pShuttle-CMV vector. This recombinant plasmid was linearized by PmeI and electronically transfected into BJ5183 cells to get the recombinant adenovirus vector Ad-Runx3. The recombinant adenovirus expressing Runx3 was infected into U251 malignant glioblastoma cells. The expression of exogenous Runx3 was observed by immonoblotting and its localization was detected by immunostaining using anti-Flag tag antibody., Results: The recombinant adenovirus expressing Runx3 with a Flag tag was constructed and infected into U251 glioblastoma cells. The exogenous Runx3 protein was detected only in the nuclei., Conclusion: The recombinant adenovirus expressing Runx3 with a Flag tag is constructed successfully, and the Runx3 protein expressed in the nuclei of infected cells. The study laid a foundation for further research of the function of Runx3 in gliocarcinogenesis.
- Published
- 2009