The RNA/DNA‐binding protein PSF relocates to cell membrane and contributes cells' sensitivity to antitumor drug, doxorubicin

Abstract

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.

The Effect of Protein FAM172A on Proliferation in HepG2 Cells and Investigation of the Possible Molecular Mechanism

“…After washing the beads, proteins were eluted from the beads in a 1x SDS sample buffer and boiled for 10 minutes at 99°C. The precipitates were separated by SDS-PAGE and stained with Coomassie Brilliant Blue as discussed [29].…”

Dual roles of DNA repair enzymes in RNA biology/post‐transcriptional control

“…As a result, HeLa cells depleted of APE1 express twofold to fivefold more c‐myc RNA in comparison to control cells . c‐myc mRNA is not the only substrate for APE1 as recent studies have demonstrated that CD44 mRNA, miRNAs, SARS‐corona virus RNA components, FGF2, and FGFR3 mRNAs are also cleaved by APE1 in vitro …”

“…13 c-myc mRNA is not the only substrate for APE1 as recent studies have demonstrated that CD44 mRNA, miR-NAs, SARS-corona virus RNA components, FGF2, and FGFR3 mRNAs are also cleaved by APE1 in vitro. [74][75][76] APE1 has also been shown to play an important role in rRNA quality control. This function was characterized through the identification of an interaction between the APE1 N-terminal region and nucleophosmin (NPM1), a nucleolar protein involved in rRNA processing and ribosome biogenesis 16 (Figure 2).…”

Long non-coding RNAs drive metastatic progression in melanoma (Review)

“…Data investigating the effects of llme23 overexpression and depletion is more limited than for HOTAIR and SPRY4-IT1, however, Llme23-depleted YUSAC cells displayed a significant decrease in their ability to form colonies in soft-agar and most significantly, these same cells displayed ~75% decrease in tumour volume at day 38 post-injection into nude mice (58). taking into consideration the emerging literature around PSf and cancer (63,64), it will be extremely interesting to observe the delineation of Llme23 function in the literature, which to date is restricted to the article discussed here.…”