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Identification of proteins responsible for the multiple drug resistance in 5-fluorouracil-induced breast cancer cell using proteomics analysis

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Abstract

Purpose

This study aimed to explore the mechanism of multi-drug resistance (MDR) in 5-fluorouracil (5-FU)-induced breast cancer cell MCF-7.

Methods

MCF-7 cells were exposed in stepwise escalating concentration of 5-FU to develop the resistant cell line, MCF-7/5-FU. Biological and molecular characteristics of the cells were studied through MTT, flow cytometry, real-time PCR, western-blot, and the global protein profiles between MCF-7/5-FU and parental MCF-7 were compared using proteomic approach. Then some of the differentially expressed proteins were validated by western-blot. In addition, the role of 14-3-3σ was validated using gene transfection.

Results

Drug resistance of MCF-7/5-FU cells to 5-FU, MX, cDDP, ADM, TAXOL all increased significantly compared with MCF-7 cells and that maybe related to BCRP, but not MDR1 and MRP1. Differentially expressed proteins between MCF-7/5-FU and MCF-7 cells were identified; 12 proteins were up-regulated and 18 proteins were down-regulated in MCF-7/5-FU cells. Expressive levels of some proteins in western-blot validation were consistent with the results in proteomic analysis. Enforced 14-3-3σ expression can increase the sensitivity of MCF-7/5-FU cells to 5-FU and cDDP.

Conclusion

MDR of MCF-7/5-FU likely associated with differentially expressed proteins and 14-3-3σ may play a positive role in chemotherapy. These findings may provide theoretical support for the prediction of chemotherapeutic response and reverse of MDR.

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Abbreviations

5-FU:

5-Fluorouracil

MX:

Mitoxantrone

cDDP:

Cisplatin

ADM:

Adramycin

BCRP:

Breast cancer resistance protein

MDR1:

P-glycoprotein

MRP1:

Multidrug resistance-associated protein

2-DE:

Two-dimensional gel electrophoresis

MALDI-TOF–MS:

Matrix-assisted laser desorption/ionization-time of flight mass spectrometry

MDR:

Multi-drug resistance

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Authors and Affiliations

  1. Cancer Research Institute, Xiangya School of Medicine, Central South University, Xiangya Road #110, Changsha, 410078, Hunan, People’s Republic of China

    Guopei Zheng, Renkui Ding, Yanhui Yu, Yongmei Ouyang, Zhuchu Chen & Zhimin He

  2. Key Laboratory of Cancer Proteomics of Ministry of Health of China, Xiangya Hospital, Central South University, 87# Xiangya Road, Changsha, 410078, Hunan, People’s Republic of China

    Fang Peng, Zhuchu Chen & Zhiqiang Xiao

Authors
  1. Guopei Zheng
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  2. Fang Peng
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  3. Renkui Ding
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  4. Yanhui Yu
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  5. Yongmei Ouyang
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  6. Zhuchu Chen
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  7. Zhiqiang Xiao
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  8. Zhimin He
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Corresponding author

Correspondence to Zhimin He.

Additional information

G. Zheng and F. Peng contributed equally to this work.

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Zheng, G., Peng, F., Ding, R. et al. Identification of proteins responsible for the multiple drug resistance in 5-fluorouracil-induced breast cancer cell using proteomics analysis. J Cancer Res Clin Oncol 136, 1477–1488 (2010). https://doi.org/10.1007/s00432-010-0805-z

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  • DOI: https://doi.org/10.1007/s00432-010-0805-z

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