%0 Journal Article
%T A novel method for sample preparation of fresh lung cancer tissue for proteomics analysis by tumor cell enrichment and removal of blood contaminants
%A Luigi De Petris
%A Maria Pernemalm
%A G£¿ran Elmberger
%A Per Bergman
%A Lotta Orre
%A Rolf Lewensohn
%A Janne Lehti£¿
%J Proteome Science
%D 2010
%I BioMed Central
%R 10.1186/1477-5956-8-9
%X Cytological analyses on cytospin specimens showed that the percentage of tumoral cells in the ETS samples ranged from 20% to 70%. In the normal lung samples the percentage of epithelial cells was less then 10%. The reproducibility of the sample preparation protocol was very good, with coefficient of variation at the peptide level and at the protein level of 13% and 7%, respectively. Proteomics analysis led to the identification of a significantly higher number of proteins in the ETS samples than in the FF samples (244 vs 109, respectively). Albumin and hemoglobin were among the top 5 most abundant proteins identified in the FF samples, showing a high contamination with blood and plasma proteins, whereas ubiquitin and the mitochondrial ATP synthase 5A1 where among the top 5 most abundant proteins in the ETS samples.The method is feasible and reproducible. We could obtain a fair enrichment of cells but the major benefit of the method was an effective removal of contaminants from red blood cells and plasma proteins resulting in larger proteome coverage compared to the direct lysis of frozen samples. This sample preparation method may be successfully implemented for the discovery of lung cancer biomarkers on tissue samples using mass spectrometry-based proteomics.Lung cancer is the number one cause of cancer-related deaths worldwide [1]. According to clinico-pathological criteria lung cancer can be divided into two major groups: small-cell lung cancer (SCLC) and non-small-cell lung cancer (NSCLC). The latter accounts for 85% of all lung cancer cases, and further comprises diverse histological subtypes, such as adenocarcinoma, squamous-cell carcinoma and undifferentiated large-cell carcinoma. All NSCLC subtypes share some common features, including a marked resistance to anticancer drugs, lack of effective screening strategies to diagnose the disease when it still is at a potentially curable early stage, and an unacceptably low overall survival, with as few as 15% of pat
%U http://www.proteomesci.com/content/8/1/9