Involvement of annexin A1 in multidrug resistance of K562/ADR cells identified by the proteomic study.
Multidrug resistance (MDR) to chemotherapy is a significant barrier to the effective treatment of chromic myeloid leukemia (CML). In an attempt to identify more factors associated with MDR for an understanding of the mechanism, we first established an adriamycin (ADR)-resistant human erythroleukemia cell line K562/ADR by stepwise selection in vitro using ADR. Besides the elevated resistance to ADR, the K562/ADR cells also showed significantly increased crossed-resistance to vincristin and Gleevec, compared to the parental K562 cells. Then we compared the global protein profiles between K562 and K562/ADR cells. Following two-dimensional gel electrophoresis and image analysis, some of the proteins with different levels between the two cell lines were identified by MALDI TOF/TOF mass spectrometry and Western blot analysis. The differentially expressed proteins were classified into groups based on their functions: calcium-binding proteins, chaperones, metabolic enzymes, proteins related to protein synthesis or DNA synthesis, and proteins related to signal transduction. In particular, ANXA1, a protein that was downregulated in K562/ADR, was analyzed further for its involvement in MDR by transfection and subsequent assays. The functional validation showed that the downregulated ANXA1 expression contributes considerably to the observed drug resistance in K562/ADR cells. These data will be valuable for further study of the mechanisms of MDR and may reveal a potential new diagnostic marker to chemotherapy.
Zhu
Fengjia
F
Sir Run Run Shaw Institute of Clinical Medicine, College of Medicine, Zhejiang University and Key Laboratory of Biotherapy of Zhejiang Province , Hangzhou, People's Republic of China.
Wang
Yi
Y
Zeng
Su
S
Fu
Xiaoyun
X
Wang
Linbo
L
Cao
Jiang
J
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D000818:Animals; D017305:Annexin A1 / Q000235:genetics / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism; D004317:Doxorubicin / Q000494:pharmacology* / Q000627:therapeutic use; D018432:Drug Resistance, Multiple / Q000502:physiology*; D006801:Humans; D020014:K562 Cells / Q000187:drug effects / Q000502:physiology; D004915:Leukemia, Erythroblastic, Acute / Q000188:drug therapy; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000188:drug therapy; D008969:Molecular Sequence Data; D040081:Protein Array Analysis; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D017305:Annexin A1; D002135:Calcium-Binding Proteins; D004317:Doxorubicin