Proteomic Profiling Identifies Pathways Dysregulated in Non-small Cell Lung Cancer and an Inverse Association of AMPK and Adhesion Pathways with Recurrence

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Proteomic Profiling Identifies Pathways Dysregulated in Non-small Cell Lung Cancer and an Inverse Association of AMPK and Adhesion Pathways with Recurrence Meera Nanjundan, PhD, Lauren Averett Byers, MD, MS, Mark S. Carey, PhD, Doris R. Siwak, PhD, Maria Gabriela Raso, MD, Lixia Diao, PhD, Jing Wang, PhD, Kevin R. Coombes, PhD, Jack A. Roth, MD, Gordon B. Mills, MD, PhD, Ignacio I. Wistuba, MD, John D. Minna, MD, John V. Heymach, MD, PhD Journal of Thoracic Oncology Volume 5, Issue 12, Pages 1894-1904 (December 2010) DOI: 10.1097/JTO.0b013e3181f2a266 Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 Unsupervised hierarchical clustering identifies distinct protein expression patterns between normal lung and non-small cell lung cancer (NSCLC) tumors. Levels of 63 proteins and phosphoproteins were determined by reverse-phase protein lysate arrays in paired normal lung (blue) and NSCLC (red) samples from 46 patients. Unsupervised hierarchical clustering separated samples into two main groups based on differences in protein expression. One group contained a majority of the tumor samples (red), whereas the other contained mostly normal lung (blue), indicating major differences in protein expression between tumor and normal lung, even within an individual patient. Replicate proteins, such as p-p38(T180), pLKB1, and cyclin D1, clustered next to each other. Journal of Thoracic Oncology 2010 5, 1894-1904DOI: (10.1097/JTO.0b013e3181f2a266) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 Identification of proteins differentially expressed between paired normal lung and non-small cell lung cancer tumor samples. A, Protein and phosphoprotein levels for 63 markers were compared between normal tissue (blue) and tumor tissue (red) from patients who underwent surgical resection of their tumors. Protein levels were compared between the two groups by t test and those with p value <0.005 (false discovery rate <1%) are shown. B, The ratio of phospho- to total protein levels are shown for Akt and FAK. Journal of Thoracic Oncology 2010 5, 1894-1904DOI: (10.1097/JTO.0b013e3181f2a266) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 A four-marker proteomic signature discriminates tumor from normal lung tissue. A, The receiver operating characteristic curve shows good performance of the model to correctly classify tumor and normal tissues. B, Two-way hierarchical clustering of the normal (blue) and tumor (red) tissues from the training set shows that these tissues are well differentiated by levels of the four markers (p70S6K, cyclin B1, pSrc(Y527), and caveolin). C, Similarly, the four markers also separate normal (blue) from tumor (red) in the test set. Journal of Thoracic Oncology 2010 5, 1894-1904DOI: (10.1097/JTO.0b013e3181f2a266) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions