Proteomic analysis of ubiquitination-associated proteins in a cisplatin-resistant human lung adenocarcinoma cell line

  • Authors:
    • Xia Qin
    • Shizhi Chen
    • Zongyin Qiu
    • Yuan Zhang
    • Feng Qiu
  • View Affiliations

  • Published online on: February 13, 2012     https://doi.org/10.3892/ijmm.2012.912
  • Pages: 791-800
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Abstract

The objective of this study was to screen for ubiquitination-associated proteins involved in cisplatin resistance in a human lung adenocarcinoma cell strain using a comparative proteomic strategy. We employed 1D SDS-PAGE to separate ubiquitinated proteins isolated and enriched from A549 and A549/CDDP lysates via affinity chromatography. The differentially expressed bands between 45-85 kDa were subsequently hydrolyzed by trypsin and subjected to HPLC-CHIP-MS/MS analysis. Of the 11 proteins identified, 7 proteins were monoubiquitinated or polyubiquitinated substrates and 4 proteins were E3 ubiquitin ligase-associated proteins. The results of western blotting and confocal laser scanning microscopy indicated that the expression levels of the E3 ubiquitin ligases RNF6, LRSAM1 and TRIM25 in A549 cells were significantly lower than those in the A549/CDDP cell line. The expression levels of the above three ubiquitin ligases in both cell lines were significantly decreased upon treatment with cis-diamminedichloroplatinum (CDDP), and the expression in the A549/CDDP cell after the treatment with CDDP decreased to a lesser extent. The expression of the substrate PKM2 in the A549 cell was higher than that in the A549/CDDP cells. Moreover, the expression of PKM2 increased in the A549 cell line and decreased in the A549/CDDP cell line upon CDDP treatment. This study suggests that drug resistance is closely correlated with changes in the ubiquitination process at the protein level in a human lung adenocarcinoma cell line.

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May 2012
Volume 29 Issue 5

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Qin X, Chen S, Qiu Z, Zhang Y and Qiu F: Proteomic analysis of ubiquitination-associated proteins in a cisplatin-resistant human lung adenocarcinoma cell line. Int J Mol Med 29: 791-800, 2012
APA
Qin, X., Chen, S., Qiu, Z., Zhang, Y., & Qiu, F. (2012). Proteomic analysis of ubiquitination-associated proteins in a cisplatin-resistant human lung adenocarcinoma cell line. International Journal of Molecular Medicine, 29, 791-800. https://doi.org/10.3892/ijmm.2012.912
MLA
Qin, X., Chen, S., Qiu, Z., Zhang, Y., Qiu, F."Proteomic analysis of ubiquitination-associated proteins in a cisplatin-resistant human lung adenocarcinoma cell line". International Journal of Molecular Medicine 29.5 (2012): 791-800.
Chicago
Qin, X., Chen, S., Qiu, Z., Zhang, Y., Qiu, F."Proteomic analysis of ubiquitination-associated proteins in a cisplatin-resistant human lung adenocarcinoma cell line". International Journal of Molecular Medicine 29, no. 5 (2012): 791-800. https://doi.org/10.3892/ijmm.2012.912