Proteomic identification of differentially expressed proteins associated with the multiple drug resistance in methotrexate-resistant human breast cancer cells

  • Authors:
    • Siying Chen
    • Jiangxia Cai
    • Weipeng Zhang
    • Xiaowei Zheng
    • Sasa Hu
    • Jun Lu
    • Jianfeng Xing
    • Yalin Dong
  • View Affiliations

  • Published online on: April 16, 2014     https://doi.org/10.3892/ijo.2014.2389
  • Pages: 448-458
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Abstract

Methotrexate (MTX), as a chemotherapeutic drug, is widely used in the therapy of several cancer types. The efficiency of drug treatment is compromised by the appearance of multidrug resistance (MDR), and the underlying molecular mechanisms remain incompletely understood. We investigated the mechanism of MDR in the MTX-induced breast cancer MCF-7 cells (MCF-7/MTX) using proteomic analysis. MCF-7 drug-sensitive cells (MCF-7/S) were exposed in progressively increasing concentrations of MTX to establish the drug-resistant cell line MCF-7/MTX. The biological characteristics of the cells were analyzed by MTT, flow cytometry, quantitative PCR, western blotting and the global protein profiles of MCF-7/MTX and MCF-7/S were compared using a proteomic approach. The resistance factor of MCF-7/MTX cells was 64, and it possessed significant MDR. Seventeen differentially expressed proteins between MCF-7/MTX and MCF-7/S cells were identified, seven proteins were upregulated and 10 proteins were downregulated in MCF-7/MTX cells. We verified that the protein levels of nucleophosmin (NPM), α-enolase (ENO1) and vimentin (VIM) were upregulated, and heterogeneous nuclear ribonucleoprotein (hnRNP C1/C2), phosphoglycerate mutase 1 (PGAM1) and proteasome subunit α type-2 (PSMA2) were downregulated in MCF-7/MTX cells. The mRNA levels of NPM, VIM, hnRNP C1/C2, PGAM1 and PSMA2 were consistent with the protein expressions, but the gene expression of ENO1 was slightly downregulated. Surprisingly, knockdown of NPM by siRNA sensitized MCF-7/MTX cells to MTX and attenuated the multidrug resistance. The proteins identified, particularly NPM provides new insights into the mechanism of MDR and is expected to become a crucial molecular target for breast cancer treatment.
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July-2014
Volume 45 Issue 1

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Chen S, Cai J, Zhang W, Zheng X, Hu S, Lu J, Xing J and Dong Y: Proteomic identification of differentially expressed proteins associated with the multiple drug resistance in methotrexate-resistant human breast cancer cells. Int J Oncol 45: 448-458, 2014
APA
Chen, S., Cai, J., Zhang, W., Zheng, X., Hu, S., Lu, J. ... Dong, Y. (2014). Proteomic identification of differentially expressed proteins associated with the multiple drug resistance in methotrexate-resistant human breast cancer cells. International Journal of Oncology, 45, 448-458. https://doi.org/10.3892/ijo.2014.2389
MLA
Chen, S., Cai, J., Zhang, W., Zheng, X., Hu, S., Lu, J., Xing, J., Dong, Y."Proteomic identification of differentially expressed proteins associated with the multiple drug resistance in methotrexate-resistant human breast cancer cells". International Journal of Oncology 45.1 (2014): 448-458.
Chicago
Chen, S., Cai, J., Zhang, W., Zheng, X., Hu, S., Lu, J., Xing, J., Dong, Y."Proteomic identification of differentially expressed proteins associated with the multiple drug resistance in methotrexate-resistant human breast cancer cells". International Journal of Oncology 45, no. 1 (2014): 448-458. https://doi.org/10.3892/ijo.2014.2389