title: [Construction of 293pT2-P210 cell line enables expression of bcr/abl to be regulated by Tet-off inducing-expression-system].
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.
authors.affiliation: Department of Hematology, PLA General Hospital, Beijing 100853, China.
authors.forename: Wen-Rong
authors.initials: WR
authors.lastname: Huang
authors.forename: Zhuo-Zhuang
authors.initials: ZZ
authors.lastname: Lu
authors.forename: Li-Sheng
authors.initials: LS
authors.lastname: Wang
authors.forename: Hua
authors.initials: H
authors.lastname: Wang
authors.forename: Hai-Feng
authors.initials: HF
authors.lastname: Duan
authors.forename: Qing-Fang
authors.initials: QF
authors.lastname: Li
authors.forename: Chun-Ji
authors.initials: CJ
authors.lastname: Gao
authors.forename: Wan-Ming
authors.initials: WM
authors.lastname: DA
mesh_terms: D001483:Base Sequence; D002461:Cell Line, Transformed / Q000166:cytology / Q000502:physiology; D002892:Chromosomes, Human, Pair 22 / Q000235:genetics; D002899:Chromosomes, Human, Pair 9 / Q000235:genetics; D016044:Fusion Proteins, bcr-abl / Q000096:biosynthesis* / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D016313:Genes, abl / Q000235:genetics; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D051562:Proto-Oncogene Proteins c-bcr / Q000235:genetics; D014162:Transfection; D014178:Translocation, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D016044:Fusion Proteins, bcr-abl; C496613:BCR protein, human; D051562:Proto-Oncogene Proteins c-bcr