Construction of 293pT2-P210 cell line enables expression of bcr/abl to be regulated by Tet-off inducing-expression-system.
- Author:
Wen-Rong HUANG
1
;
Zhuo-Zhuang LU
;
Li-Sheng WANG
;
Hua WANG
;
Hai-Feng DUAN
;
Qing-Fang LI
;
Chun-Ji GAO
;
Wan-Ming DA
Author Information
1. Department of Hematology, PLA General Hospital, Beijing 100853, China.
- Publication Type:Journal Article
- MeSH:
Base Sequence;
Cell Line, Transformed;
cytology;
physiology;
Chromosomes, Human, Pair 22;
genetics;
Chromosomes, Human, Pair 9;
genetics;
Fusion Proteins, bcr-abl;
biosynthesis;
genetics;
Gene Expression Regulation, Neoplastic;
Genes, abl;
genetics;
Humans;
Leukemia, Myelogenous, Chronic, BCR-ABL Positive;
genetics;
metabolism;
pathology;
Models, Genetic;
Molecular Sequence Data;
Proto-Oncogene Proteins c-bcr;
genetics;
Transfection;
Translocation, Genetic;
Tumor Cells, Cultured
- From:
Journal of Experimental Hematology
2007;15(2):224-228
- CountryChina
- Language:Chinese
-
Abstract:
Chronic myelogenous leukemia (CML) is a clonal myeloproliferative disease of transformed hematopoietic progenitor cells. It is now clear that the chimeric bcr/abl P210(bcr/abl) fusion protein, which is generated by the reciprocal translocation t (9; 22), inhibits apoptosis and increase proliferation. P210(bcr/abl) plays a central role in the pathophysiology of CML. The purpose of this study was to construct a cell line model that bcr/abl expression can be regulated by Tet-off inducing-expression-system. The full-length b3a2 bcr/abl cDNA was subcloned into the pTRE2hyg expression vector to construct the pT2-P210 plasmid. 293 cells were firstly transfected with Tet-off plasmid and the clone that the Tet-off system can work effectively after transfected with pTRE2hyg-LUC was selected by luciferase activity assay. The pT2-P210 plasmid was then transfected into the selected clone and cells were then selected for hygromycin B and G418 resistance. The results showed that individual subclones expressing bcr/abl after withdrawing doxycycline were 293pT2-P210 cell line. In conclusion, selected 293pT2-P210 cells are cells that bcr/abl expression can be regulated by Tet-off inducing-expression-system. They are suitable thoroughly to study the function of bcr/abl fusion gene and its signal regulation mechanism.
