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The RNA/DNA‐binding protein PSF relocates to cell membrane and contributes cells' sensitivity to antitumor drug, doxorubicin
Author(s) -
Ren Simei,
She Ming,
Li Min,
Zhou Qi,
Liu Rong,
Lu Hong,
Yang Chunzheng,
Xiong Dongsheng
Publication year - 2014
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.22423
Subject(s) - cell , biology , flow cytometry , hl60 , cell culture , microbiology and biotechnology , cancer research , biochemistry , genetics
Cell surface proteins play an important role in multidrug resistance (MDR). However, the identification involving chemoresistant features for cell surface proteins is a challenge. To identify potential cell membrane markers in hematologic cancer MDR, we used a cell‐ and antibody‐based strategy of subtractive immunization coupled with cell surface comparative screening of leukemia cell lines from sensitive HL60 and resistant HL60/DOX cells. Fifty one antibodies that recognized the cell surface proteins expressed differently between the two cell lines were generated. One of them, the McAb‐5D12 not only recognizes its antigen but also block its function. Comparative analysis of immunofluorescence, flow cytometry, and mass spectrum analysis validated that the membrane antigen of McAb‐5D12 is a nucleoprotein—polypyrimidine tract binding protein associated splicing factor, PSF. Our results identified that PSF overexpressed on the membrane of sensitive cells compared with resistant cells and its relocation from the nuclear to the cell surface was common in hematological malignancy cell lines and marrow of leukemia patients. Furthermore, we found that cell surface PSF contributed to cell sensitivity by inhibiting cell proliferation. The results represent a novel and potentially useful biomarker for MDR prediction. The strategy enables the correlation of expression levels and functions of cell surface protein with some cell‐drug response traits by using antibodies. © 2013 International Society for Advancement of Cytometry