K562 cell line resistance to nilotinib induced in vitro and preliminary investigation of its mechanisms.
- Author:
Ji-shi WANG
1
;
Chang YANG
;
Qin FANG
;
Si-xi WEI
;
Cheng CHEN
;
Yuan YANG
;
Ya-ting WANG
;
Xiu-ying HU
;
Dan MA
Author Information
- Publication Type:Journal Article
- MeSH: ATP Binding Cassette Transporter, Sub-Family B; ATP-Binding Cassette, Sub-Family B, Member 1; metabolism; Caspase 3; metabolism; Drug Resistance, Neoplasm; Fusion Proteins, bcr-abl; metabolism; Gene Expression Regulation, Leukemic; Heme Oxygenase-1; metabolism; Humans; K562 Cells; drug effects; Pyrimidines; pharmacology
- From: Chinese Journal of Hematology 2012;33(11):906-910
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a bcr-abl(+) cell line resistance to nilotinib, and to investigate the possible mechanisms of resistance.
METHODSK562 cells were treated with gradually increasing concentrations of nilotinib to generate resistance cell line K562-RN. The folder of drug-resistance was evaluated by MTT assay. Cells apoptosis rate was detected by flow cytometry, the mRNA level of bcr-abl fusion gene by FISH, and the expression of apoptosis relative gene mRNA and protein (such as bcr-abl, HO-1, mdr1, Bcl-2 and caspase-3) by RQ-PCR and western blot.
RESULTSThe resistant cell line K562-RN was successfully established, with 2.01 fold resistant to nilotinib compared with K562 cell line \[the IC(50) value of nilotinib to K562 and K562-RN were (12.320 ± 1.720) µmol/L and (24.742 ± 2.310) µmol/L, respectively\]. It also had the cross resistance to adriamycin, homoharringtonine, etoposide and imatinib. Treated with different concentrations of nilotinib, cell apoptosis rate of K562-RN was significantly lower than that of K562 cells. The rate of bcr-abl gene positive cells was 92% in K562-RN by FISH assay. The mRNA and protein levels of bcr-abl, HO-1 and mdr1 expression up-regulated in K562-RN cells, while those of caspase-3 expression down-regulated, being significantly statistical difference when compared with K562 cells (P < 0.05).
CONCLUSIONHuman leukemic cell line resistance to nilotinib, K562-RN is established successfully by gradually increasing concentrations of drug. The mechanisms of resistance in K562-RN is probably associated with increasing expression of bcr-abl, HO-1, mdr1 and decreasing expression of caspase-3 mRNA and protein levels.
