JIOMICS
JIOMICS Internacional
JIOMICS Internacional
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<strong>JIOMICS</strong> | VOL 5 | ISSUE 2 | DECEMBER 2015 | 1-62<br />
JOURNAL OF INTEGRATED OMICS<br />
Journal of Integrated Omics<br />
A METHODOLOGICAL JOURNAL<br />
HTTP://WWW.<strong>JIOMICS</strong>.COM<br />
Special Issue: Proceeding Abstracts of the 4 th International Congress on Analytical Proteomics (ICAP 2015)<br />
Western blotting - detection of a specific protein in MCF7 and MCF7-Z<br />
tissue<br />
J. Ner-Kluza* 1 , A. Drabik 1 , Michael Kubbutat 2 , Andreas Lingnau 2 , P. Suder 1,5 , G. Schroeder 4 , J. Silberring 1,3<br />
1<br />
Department of Biochemistry and Neurobiology., Faculty of Materials Sciences and Ceramics. AGH University of Science and Technology.<br />
Mickiewicza 30 Ave. 3 0-059 Krakow. Poland. 2 ProQinase GmbH. Freiburg, Germany. 3 Center of Polymer and Carbon Materials. Polish<br />
Academy of Sciences. Marii Skłodowskiej-Curie 34., 41-819 Zabrze. Poland. 4 Department of Supramolecular Chemistry. Faculty of Chemistry.<br />
Adam Mickiewicz University. Umultowska 89b. 61-614 Poznan. Poland. 5 AGH University of Science and Technology. Academic Centre for<br />
Materials and Nanotechnology. Mickiewicza 30. 30–059, Kraków, Poland. *Corresponding author: nerjoanna@gmail.com<br />
Available Online: 31 December 2015<br />
Abstract<br />
Purpose: In the contemporary world, breast cancer is one of the most common and dangerous types of cancer in females. Comparison<br />
between two different variants of MCF-7 (MCF-7 vs MCF-7-Z) human breast cancer cell lines used in subcutaneous xenograft tumor models,<br />
has shown different tumor growth characteristics in mice. Glycosylated proteins are among post-translational modifications one of the most<br />
important types of proteins involved in tumor progression. The aim of the research was to search for the presence of selected glycoproteins,<br />
based on quantitative and qualitative analysis in cancer progression.<br />
Experimental description: For samples preparation we used the strategy utilizing western blotting and nanoLC-MS/MS analysis. After<br />
isolation of proteins from the tissue, glycoproteins were isolated by lectin affinity chromatography. Glycoproteins fractions were separated by<br />
the sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE), followed by two complementary analyses. One part of the gel was transferred<br />
onto PVDF membrane, and another part was CBB stained. Based on the images obtained after immunostaining, the bands were excised from<br />
the CBB-stained gel and analyzed by tandem mass spectrometry coupled to liquid chromatography (LC-MS/MS).<br />
Results: Using LC-MS/MS technique we have succeeded to identify 3000 differentially expressed proteins between MCF7 and MCF7-Z. The<br />
selected proteins were associated with cancer metastasis, which plays a vital role in tumor progression, and are involved in cell structure organization.<br />
The presence of the selected proteins was confirmed using western blotting.<br />
Conclusions: We have identified the potential biomarkers, which can be important in cancer progression.<br />
Keywords: cell lines MCF-7, glycoproteins, tumor, breast cancer.<br />
Acknowledgements: The research was supported by the grant EuroNanoMed "META" 05/EuroNanoMed/2012.<br />
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