Generation and Sequencing of Pulmonary Carcinoid Tumor Cell Lines

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Presentation transcript:

Generation and Sequencing of Pulmonary Carcinoid Tumor Cell Lines Michael K. Asiedu, PhD, Charles F. Thomas, MD, Sandra C. Tomaszek, MD, Tobias Peikert, MD, Bharati Sanyal, PhD, Shari L. Sutor, MS, Marie-Christine Aubry, MD, Peter Li, PhD, Dennis A. Wigle Journal of Thoracic Oncology Volume 9, Issue 12, Pages 1763-1771 (December 2014) DOI: 10.1097/JTO.0000000000000339 Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 Histological staining of carcinoid tumors. Photomicrograph of H&E, CGA, and SYP staining of carcinoid tumors used for establishing TC1, TC2, and TC3 cell lines. Journal of Thoracic Oncology 2014 9, 1763-1771DOI: (10.1097/JTO.0000000000000339) Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 Expression of neuroendocrine markers. A, Immunohistochemistry staining demonstrating expression of CGA (red) and SYP (green) in carcinoid cell lines. B, Western blot demonstrating CGA (75 kDa) and SYP (43 kDa) expression in the established cell lines TC1, TC2, and TC3. Journal of Thoracic Oncology 2014 9, 1763-1771DOI: (10.1097/JTO.0000000000000339) Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 Typical carcinoid tumor cell lines in vitro growth characteristics. TC1, TC2, and TC3 cells were grown in vitro with or without 10% serum and proliferation was measured at various time points. Data shown is the mean fluorescence from the proliferation assay of triplicate samples +/- SEM. Journal of Thoracic Oncology 2014 9, 1763-1771DOI: (10.1097/JTO.0000000000000339) Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 4 Colony formation of lung carcinoid tumor cells in soft agar. TC1, TC2, and TC3 cells were plated in triplicate with control NSCLC A549 cells and nonneoplastic normal human bronchial epithelial cells (NHBEC) in soft agar. After 2 weeks of culture colonies greater than 50 microns in diameter were counted. Data shown is the mean of triplicate plates +/- SEM. Journal of Thoracic Oncology 2014 9, 1763-1771DOI: (10.1097/JTO.0000000000000339) Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 5 Pulmonary carcinoid xenograft tumor. A, Gross lung specimen from xenograft mouse showing large tumor nodule (arrow). B, Xenograft tumor (H&E, 10× and 40×) with comparison to the original patient tumor (H&E, 10× and 40×) demonstrating typical neuroendocrine features in both samples. Journal of Thoracic Oncology 2014 9, 1763-1771DOI: (10.1097/JTO.0000000000000339) Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 6 Venn diagram showing common and distinct mutations in the TC3 tumor and cell line. Journal of Thoracic Oncology 2014 9, 1763-1771DOI: (10.1097/JTO.0000000000000339) Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions