Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
. 2013 Mar;139(3):509-19.
doi: 10.1007/s00432-012-1350-8. Epub 2012 Nov 27.

Identification of proteins differentially expressed in adriamycin-resistant (pumc-91/ADM) and parental (pumc-91) human bladder cancer cell lines by proteome analysis

Qian Meng  1 , Ting Lei, Min Zhang, Jin Zhao, Xu-Hong Zhao, Man Zhang
Affiliations
Comparative Study

Identification of proteins differentially expressed in adriamycin-resistant (pumc-91/ADM) and parental (pumc-91) human bladder cancer cell lines by proteome analysis

Qian Meng et al. J Cancer Res Clin Oncol. 2013 Mar.

Abstract

Purpose: Resistance to chemotherapy drugs remains a difficult problem in bladder cancer treatment. Protein expression is an important factor underlying multidrug resistance (MDR) in bladder cancer. The aim of the study was to explore differentially expressed proteins responsible for MDR between an adriamycin-resistant human bladder cancer cell line (pumc-91/ADM) and its parental cell line (pumc-91).

Methods: Two-dimensional gel electrophoresis (2-DE) combining image analysis was used to screen the differentially expressed protein spots between the pumc-91/ADM and pumc-91 cell lines. Then, the protein spots were identified using MALDI-TOF/TOF mass spectrometry. Among the identified proteins, annexin A2 (ANXA2) and nucleophosmin (NPM1) were then further verified using RT-PCR and Western blot analysis.

Results: A total of 30 proteins, including 19 up-regulated and 11 down-regulated proteins, were successfully identified in pumc-91/ADM. According to their different functions, these 30 proteins were classified into 12 categories. Annexin A2 (ANXA2) and nucleophosmin (NPM1) were up-regulated in pumc-91/ADM compared with pumc-91.

Conclusion: The proteins identified may have an important clinical significance in MDR, and ANXA2 and NPM1 may take part in mechanism of MDR in bladder cancer.

PubMed Disclaimer

Conflict of interest statement

We declare that we have no conflict of interest.

Figures

Fig. 1
Fig. 1
Representative two-dimensional electrophoresis gels (pI 3–10, 24 cm) showed that 34 proteins of differential expression (signed the numbers) between the drug-resistant cell line pumc-91/ADM and its parental cell line pumc-91
Fig. 2
Fig. 2
Through MALDI-TOF/TOF analysis, the protein spots 11 and 2 were identified as ANXA2 and NPM1. The peptide mass fingerprinting (PMF) of ANXA2 and NPM1 is showed in (a, b). The matched amino acid sequences (marked with red) of ANXA2 and NPM1 are showed in (c, d)
Fig. 2
Fig. 2
Through MALDI-TOF/TOF analysis, the protein spots 11 and 2 were identified as ANXA2 and NPM1. The peptide mass fingerprinting (PMF) of ANXA2 and NPM1 is showed in (a, b). The matched amino acid sequences (marked with red) of ANXA2 and NPM1 are showed in (c, d)
Fig. 3
Fig. 3
A representative MS/MS spectrum of ANXA2 (m/z = 1,086.4807) and NPM1 (m/z = 2,243.2314), respectively, is showed in (a, b). The identified peptide sequence was AYTNFDAER and MSVQPTVSLGGFEITPPVVLR
Fig. 4
Fig. 4
Pie chart showed the 13 functional categories as a percentage of the differentially expressed proteins
Fig. 5
Fig. 5
a, c showed the results of ANXA2 and NPM1 PCR products. b, d The relative mRNA expression levels of them between the pumc-91/ADM and pumc-91 cell lines as verified by RT-PCR, after normalization against β-actin. RT-PCR analysis was repeated at least three times
Fig. 6
Fig. 6
ANXA2 (Protein Spot 11) and NPM1 (Protein Spot 2) in the pumc-91/ADM and pumc-91 cell lines was verified by Western blot
See this image and copyright information in PMC

References

    1. Baguley BC (2010) Multi-drug resistance in cancer. Methods Mol Biol 596:1–9 - PubMed
    1. Borer RA, Lehner CF, Eppenberger HM et al (1989) Major nucleolar proteins shuttle between nucleus and cytoplasm. Cell 56:379–390 - PubMed
    1. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254 - PubMed
    1. Cole SP, Pinkoski MJ, Bhardwaj G et al (1992) Elevated expression of annexin II (lipocortin II, p36) in a multidrug resistant small cell lung cancer cell line. Br J Cancer 65(4):498–502 - PMC - PubMed
    1. Colombo E, Marine JC, Danovi D et al (2002) Nucleophosmin regulates the stability and transcriptional activity of p53. Nat Cell Biol 4:529–533 - PubMed

Publication types

MeSH terms

LinkOut - more resources