Use of MicroRNA Expression Levels to Predict Outcomes in Resected Stage I Non- small Cell Lung Cancer Eric Duncavage, MD, Boone Goodgame, MD, Ananth Sezhiyan,

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Use of MicroRNA Expression Levels to Predict Outcomes in Resected Stage I Non- small Cell Lung Cancer Eric Duncavage, MD, Boone Goodgame, MD, Ananth Sezhiyan, BS, Ramaswamy Govindan, MD, John Pfeifer, MD, PhD Journal of Thoracic Oncology Volume 5, Issue 11, Pages 1755-1763 (November 2010) DOI: 10.1097/JTO.0b013e3181f3909d Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 Cases of stage I non-small cell carcinoma included predominantly squamous cell carcinomas (A) and adenocarcinomas of the lung (B) (×100 original magnification). RNA was extracted from areas of tumor that contained greater than 70% tumor nuclei and minimal necrosis. Journal of Thoracic Oncology 2010 5, 1755-1763DOI: (10.1097/JTO.0b013e3181f3909d) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 Expression of averaged triplicate microRNA (miRNA) data normalized to rnu-6b (log plots). Journal of Thoracic Oncology 2010 5, 1755-1763DOI: (10.1097/JTO.0b013e3181f3909d) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 Kaplan-Meier freedom-from-recurrence curves for miR-221 expression. miR-221 values were first dichotomized about a mean expression value and those cases with expression above the mean value designated “high expression” and those below designated “low expression.” Journal of Thoracic Oncology 2010 5, 1755-1763DOI: (10.1097/JTO.0b013e3181f3909d) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 4 MiR-221 expression by recurrence status in adenocarcinoma (A) and squamous cell carcinoma (B). Journal of Thoracic Oncology 2010 5, 1755-1763DOI: (10.1097/JTO.0b013e3181f3909d) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 5 A comparison of microRNA (miRNA) expression in tumor (red) and paired, adjacent normal lung (blue) for both nonrecurrent tumors (left panel) and recurrent tumors (right panel). Journal of Thoracic Oncology 2010 5, 1755-1763DOI: (10.1097/JTO.0b013e3181f3909d) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 6 Expression of selected miR-221 target genes. Target gene expression was normalized to a multiplexed beta-actin control. Journal of Thoracic Oncology 2010 5, 1755-1763DOI: (10.1097/JTO.0b013e3181f3909d) Copyright © 2010 International Association for the Study of Lung Cancer Terms and Conditions