Identification of Chemoresistance-Related Cell-Surface Glycoproteins in Leukemia Cells and Functional Validation of Candidate Glycoproteins

Abstract

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.

FK506 Binding Protein 10 Is Overexpressed and Promotes Renal Cell Carcinoma

“…Different from all the other family proteins, FKBP10 contains 4 PPIase domains [20,21] . FKBP10 has been implicated in ovarian cancer [17,22] , colorectal cancer [23] , KRAS-induced lung cancer [18] , prostate cancer [24], and leukemia [25] .…”

CLPTM1L/CRR9 ectodomain interaction with GRP78 at the cell surface signals for survival and chemoresistance upon ER stress in pancreatic adenocarcinoma cells

“…Our working model, supported by the findings herein, is that ER stress and UPR caused by microenvironmental, oncogenic, or therapeutic stress can result in increased GRP78 accumulation and translocation of both GRP78 and CLPTM1L to the cell surface. N‐glycosylated CLPTM1L peptides have been identified on pluripotent stem cells and chemoresistant human leukemia cells by mass spectrometry after surface‐protein‐capture by biotinylation . Interestingly, CLPTM1L was detected as a surface protein only in AraC resistant, but not sensitive human chronic myelogenous leukemia cells …”

“…N‐glycosylated CLPTM1L peptides have been identified on pluripotent stem cells and chemoresistant human leukemia cells by mass spectrometry after surface‐protein‐capture by biotinylation . Interestingly, CLPTM1L was detected as a surface protein only in AraC resistant, but not sensitive human chronic myelogenous leukemia cells …”

Use of monolithic supports for high‐throughput protein and peptide separation in proteomics

“…This system provided quantification of about 1000 proteins in 30 h from trypsinized liver cell lysate. Similar procedure applied for analysis of leukemia cell lines on the same MS system operated in top 6 DDA mode quantified more than 1200 proteins .…”