Expression and localization of adenovirus-mediated transcriptional factor Runx3 in malignant glioblastoma cells
10.3321/j.issn:1007-8738.2009.10.002
- VernacularTitle:重组腺病毒介导的转录因子Runx3在神经胶质瘤细胞中的表达及其亚细胞定位
- Author:
Wendong BAI
;
Rui ZHANG
;
Yanling MENG
;
Yunxin CAO
;
Tao WANG
;
Jing ZHAO
;
Angang YANG
- Publication Type:Journal Article
- Keywords:
Runx3;
adenovirus;
malignant glioma cells
- From:
Journal of Cellular and Molecular Immunology
2009;25(10):866-869
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct the replicative defecient 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-CMVRunx3 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 Pmel 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.
