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Comparative proteomic analysis of paclitaxel sensitive A549 lung adenocarcinoma cell line and its resistant counterpart A549-Taxol

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Abstract

Purpose

Paclitaxel is used as the first-line chemotherapy for Non-Small Cell Lung Cancer (NSCLC), but acquired resistance becomes a critical problem. Several mechanisms have been proposed in paclitaxel resistance, but they are not sufficient to exhaustively explain this resistance emergence. To better investigate molecular resistance mechanisms, a comparative proteomic approach was carried out to identify differentially expressed proteins between human lung adenocarcinoma A549 cell line (paclitaxel sensitive) and A549-Taxol cell line (acquired resistant).

Methods

A paclitaxel-resistant subline (A549-Taxol) derived from the parental-sensitive cell line A549 was established by stepwise selection by paclitaxel. Total proteins in the two cell lines were separated by fluorescent differential gel electrophoresis (DIGE). Image analysis was carried out with the DeCyder 2D 6.5 software. Proteins associated with chemoresistance process were identified by matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF–MS/MS). Some key molecules were valuated by Western blot.

Results

Thirty proteins were identified and grouped into eight main functional classes according to the biological processes in which they are likely to participate, i.e. signal transduction, cytoskeleton, redox reaction, energy and metabolism, and so on. Alterations of these processes might be involved in paclitaxel resistance. Most of the proteins showed mitochondrial and cytoplasm location. The up-regulation of CK8, CK18, ALDH1, CAST and ANX I in A549-Taxol cell line was verified by Western blot, in coincidence with the data obtained from proteomic analysis.

Conclusion

For the first time, differentially expressed proteins between paclitaxel-sensitive cell line and paclitaxel-resistant one were explored by comparative proteomic approach in human lung adenocarcinoma. It may be useful for further studying of resistance mechanisms and screening of resistance biomarkers, so as to develop tailored therapeutic strategies.

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Abbreviations

2-DE:

Two-dimensional gel electrophoresis

CK8:

Cytokeratin-8

CK18:

Cytokeratin-18

G6PD:

Glucose-6-phosphate 1-dehydrogenase

RALDH1:

Retinal dehydrogenase 1

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Acknowledgments

This work was supported by a grant from the Shanghai United Municipal Hospitals Project (to Hui-fang SHA) (NO.SHDC12007103), Shanghai Municipal Natural Science Foundation (to Qiang-ling SUN) (NO.10ZR1428100), and grant for cancer research from Shanghai charity foundation (to Qiang-ling SUN). We wish to thank all the researchers of public service platform of Shanghai Shenkang medical center which located in Ruijin Hospital for their assistance with the mass spectrometric analyses in this study.

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Authors and Affiliations

  1. Basic Research Laboratory, Shanghai Chest Hospital Affiliated to Shanghai Jiaotong University, Shanghai, 200030, China

    Qiang-ling Sun, Hui-fang Sha, Xiao-hua Yang & Guo-liang Bao

  2. Institute of Hematology, Ruijin Hospital of Shanghai Jiaotong University, Shanghai, 200025, China

    Jing Lu & Yin-yin Xie

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  1. Qiang-ling Sun
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  2. Hui-fang Sha
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  3. Xiao-hua Yang
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  4. Guo-liang Bao
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  5. Jing Lu
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Correspondence to Hui-fang Sha.

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Sun, Ql., Sha, Hf., Yang, Xh. et al. Comparative proteomic analysis of paclitaxel sensitive A549 lung adenocarcinoma cell line and its resistant counterpart A549-Taxol. J Cancer Res Clin Oncol 137, 521–532 (2011). https://doi.org/10.1007/s00432-010-0913-9

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  • DOI: https://doi.org/10.1007/s00432-010-0913-9

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