Identification of chemoresistance-related cell-surface glycoproteins in leukemia cells and functional validation of candidate glycoproteins.
Chemoresistance remains the most significant obstacle to successful chemotherapy for leukemia, and its exact mechanism is still unknown. In this work, we used the cell-surface capturing method together with quantitative proteomics to investigate differences in the glycoproteomes of adriamycin-sensitive and adriamycin-resistant leukemia cells. Two quantitative methods, isotopic dimethyl labeling and SWATH, were used to quantify glycoproteins, and 35 glycoproteins were quantified by both methods. High correlation was observed between the glycoproteins quantified by the above two methods, and 15 glycoproteins displayed a consistent significant change trend in both sets of quantitative results. These 15 proteins included classical multidrug resistance-related glycoproteins such as ABCB1 as well as a set of novel glycoproteins that have not previously been reported to be associated with chemoresistance in leukemia cells. Further validation with quantitative real-time PCR and Western blotting confirmed the proteomic screening results. Subsequent functional experiments based on RNA interference technology showed that CTSD, FKBP10, and SLC2A1 are novel genes that participate in the acquisition and maintenance of the adriamycin-resistant phenotype in leukemia cells.
Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , 457 Zhongshan Road, Dalian 116023, China.
Zhen
Z
Sun
Jiaqiang
J
Dong
Song
S
Zhang
Zhengyan
Z
Hu
Kai
K
Cheng
Kai
K
Li
Bo
B
Xu
Mingliang
M
Ye
Yongzhan
Y
Nie
Daiming
D
Fan
Hanfa
H
Zou
D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D002402:Cathepsin D / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D004317:Doxorubicin / Q000494:pharmacology; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D015973:Gene Expression Regulation, Leukemic; D051272:Glucose Transporter Type 1 / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D040901:Proteomics / Q000379:methods; D012984:Software; D013194:Staining and Labeling / Q000379:methods; D022021:Tacrolimus Binding Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry
D000903:Antibiotics, Antineoplastic; D051272:Glucose Transporter Type 1; D008562:Membrane Glycoproteins; C495677:SLC2A1 protein, human; D004317:Doxorubicin; C533922:CTSD protein, human; D002402:Cathepsin D; D022021:Tacrolimus Binding Proteins; C110054:FKBP10 protein, human