K562 cell line resistance to nilotinib induced in vitro and preliminary investigation of its mechanisms / 中华血液学杂志
Chinese Journal of Hematology
; (12): 906-910, 2012.
Article
in Zh
| WPRIM
| ID: wpr-278303
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To establish a bcr-abl(+) cell line resistance to nilotinib, and to investigate the possible mechanisms of resistance.</p><p><b>METHODS</b>K562 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.</p><p><b>RESULTS</b>The 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).</p><p><b>CONCLUSION</b>Human 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.</p>
Full text:
1
Index:
WPRIM
Main subject:
Pharmacology
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Pyrimidines
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Gene Expression Regulation, Leukemic
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Fusion Proteins, bcr-abl
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ATP Binding Cassette Transporter, Subfamily B, Member 1
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Drug Resistance, Neoplasm
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ATP Binding Cassette Transporter, Subfamily B
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K562 Cells
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Heme Oxygenase-1
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Caspase 3
Limits:
Humans
Language:
Zh
Journal:
Chinese Journal of Hematology
Year:
2012
Type:
Article