中国药理学与毒理学杂志2012，Vol.26Issue(4)：563-569,7.DOI:10.3867/j.issn.1000-3002.2012.04.017
酪氨酸激酶抑制剂伊马替尼耐药K562细胞系的建立及其耐药特征
Establishment of human K562 cell line resistant to tyrosine kinase inhibitor imatinib and its resistance characters
摘要
Abstract
OBJECTIVE To screen imatinib(STI-571) resistant human K562 cell line and to study the imatinib resistance mechanisms. METHODS Human K562 cells were induced by culture in the medium with gradually increasing imatinib concentrations to establish the imatinib-resist-ant K562 (K562R) cells. Biological and pharmacological characters of the cells were examined by MTT method, and cell phenotype and apoptosis were determined. The expression of B cell receptor-c-Abelson (BCR-ABL) , multi-drug resistance (MDR) and p-glycoprotein(p-GP) genes was examined by semi-QPCR and Western blotting. RESULTS Compared to K562 cells ( IC50 = 0.01 μmol·L-1), K562R cells exhibited 235.0 fold resistance to imatinib (IC50 =2.35 μmol· L-1). Besides, K562R also became resistant to homoharringtonine ( 13. 2 folds) , vincristine (63. 2 folds) and daunorubicin ( 11.8 folds ). These resistance abilities consisted with the expression of BCR-ABL, MDR and p-GP genes. PCR results showed that the expression of BCR-ABL, MDR and p-GP genes increased in, K562R cells compared with K562 cells. The DNA sequencing analysis showed that a A→C point-mutant occurred in the BCR-ABL gene sequence of K562R cells, and as-paragine was submitted by lysine in the 231st site of kinase-dornain. CONCLUSION K562R cells are successfully constructed. The expression of BCR-ABL, MDR and p-GP genes is up-regulated in K562R cells. There is a point-mutant in BCR-ABL gene of K562R cells.关键词
伊马替尼/酪氨酸激酶抑制剂/K562细胞/抗药性,肿瘤Key words
imatinib/tyrosine kinase inhibitor/K562 cells/drug resistance, neoplasm分类
医药卫生引用本文复制引用
冯帆,张琼,刘洪英,李松,王莉莉..酪氨酸激酶抑制剂伊马替尼耐药K562细胞系的建立及其耐药特征[J].中国药理学与毒理学杂志,2012,26(4):563-569,7.基金项目
国家自然科学基金(30800323) （30800323）
国家"重大新药创制"科技重大专项(2009ZX09501-031) （2009ZX09501-031）
