N‑Glycoprotein Analysis Discovers New Up-Regulated Glycoproteins in Colorectal Cancer Tissue

Colorectal cancer is one of the leading causes of death due to cancer worldwide. Therefore, the identification of high-specificity and -sensitivity biomarkers for the early detection of colorectal cancer is urgently needed. Post-translational modifications, such as glycosylation, are known to play an important role in cancer progression. In the present work, we used a quantitative proteomic technique based on ¹⁸O stable isotope labeling to identify differentially expressed N-linked glycoproteins in colorectal cancer tissue samples compared with healthy colorectal tissue from 19 patients undergoing colorectal cancer surgery. We identified 54 up-regulated glycoproteins in colorectal cancer samples, therefore potentially involved in the biological processes of tumorigenesis. In particular, nine of these (PLOD2, DPEP1, SE1L1, CD82, PAR1, PLOD3, S12A2, LAMP3, OLFM4) were found to be up-regulated in the great majority of the cohort, and, interestingly, the association with colorectal cancer of four (PLOD2, S12A2, PLOD3, CD82) has not been hitherto described

Adjacent normal mucosa section and colon cancer section immunostained for the evaluation of MCDPT and VEGF expression at x400 magnification respectively.

<p>In <b>A</b> a primary anti-tryptase antibody has been employed. Arrow indicates a single and unique red mast cells positive to tryptase in the observed field. In <b>B</b> a primary anti-VEGF antibody has been employed. Many red immunostained cancer cells. Big arrows indicate single red cytoplasmic cancer cells positive to VEGF.</p

Colon cancer sections immunostained for the detail evaluation of MCDPT and MVD at x1000 magnification in oil respectively.

<p>In <b>A</b> a primary anti-tryptase antibody has been employed. The big arrows indicate single red mast cells positive to tryptase. The small arrow indicates the degranulation front of the mast cell forming a microvessel. In <b>B</b> a primary anti CD-34 antibody has been employed. The big arrow indicates a red positive microvessel. The small arrow indicates the nucleus of the endothelial cell. Note the lumen of the microvessel.</p

Colon cancer sections immunostained for the evaluation of MCDPT and MVD at x400 magnification, high MCDPT and MVD respectively.

<p>In <b>A</b> a primary anti-tryptase antibody has been employed. Arrows indicate red mast cells positive to tryptase. In <b>B</b> a primary anti CD-34 antibody has been employed. Arrows indicate red positive microvessels.</p

Clinico-pathological features of patients.

<p>Clinico-pathological features of patients.</p

Correlation analysis between MCDPT and STLBS.

<p>Correlation analysis between MCDPT and STLBS.</p

Additional file 5: of Next-generation sequencing analysis of receptor-type tyrosine kinase genes in surgically resected colon cancer: identification of gain-of-function mutations in the RET proto-oncogene

Table S5. List of mutations identified in CC20 after the next generation sequencing analysis pipeline. (XLSX 17 kb

Differences between STLBS (red line) and STLAS (blu line).

<p>Differences between STLBS (red line) and STLAS (blu line).</p

Additional file 3: of Next-generation sequencing analysis of receptor-type tyrosine kinase genes in surgically resected colon cancer: identification of gain-of-function mutations in the RET proto-oncogene

Table S3. List of mutated RTKs in each patient included in the study. (XLSX 11 kb

Additional file 5: of Next-generation sequencing analysis of receptor-type tyrosine kinase genes in surgically resected colon cancer: identification of gain-of-function mutations in the RET proto-oncogene

Table S5. List of mutations identified in CC20 after the next generation sequencing analysis pipeline. (XLSX 17 kb