MiR-205 and MiR-375 MicroRNA Assays to Distinguish Squamous Cell Carcinoma from Adenocarcinoma in Lung Cancer Biopsies Santosh Patnaik, MD, PhD, Reema.

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MiR-205 and MiR-375 MicroRNA Assays to Distinguish Squamous Cell Carcinoma from Adenocarcinoma in Lung Cancer Biopsies Santosh Patnaik, MD, PhD, Reema Mallick, MD, Eric Kannisto, MS, Rohit Sharma, MD, Wiam Bshara, MD, Sai Yendamuri, MD, Samjot Singh Dhillon, MD Journal of Thoracic Oncology Volume 10, Issue 3, Pages 446-453 (March 2015) DOI: 10.1097/JTO.0000000000000423 Copyright © 2015 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 Histotypic microRNAs to distinguish lung SCC and AC identified in analyses of data from the Cancer Genome Atlas. Count per million microRNAs in primary AC (n = 334, black) and SCC (n = 300, gray) tumor or tumor-adjacent normal lung tissues (n = 79, white) for 10 of the 12 microRNAs identified in the analyses are depicted as mean and 95% confidence interval values. SCC, squamous cell carcinoma; AC, adenocarcinoma. Journal of Thoracic Oncology 2015 10, 446-453DOI: (10.1097/JTO.0000000000000423) Copyright © 2015 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 Correlation between small RNA quantifications obtained by RT-PCR using two amounts of tissue RNA. Measurements obtained using 0.2 ng RNA of 14 resected lung cancers of U6B and miR-21, miR-205, and miR-375 as quantification cycle (Cq) values, and of miR-205 relative to U6B and miR-21 (Lebanony score), or only U6B (miR-205 ΔCqU6B) are plotted against those obtained with 5 ng RNA. The Pearson correlation coefficient (r) with 95% confidence interval, and the linear regression line generated with the least squares fitting technique are also shown. RT-PCR, reverse transcription-polymerase chain reaction. Journal of Thoracic Oncology 2015 10, 446-453DOI: (10.1097/JTO.0000000000000423) Copyright © 2015 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 Expression of miR-205 in lung SCC and AC resectates. RT-PCR was used to quantify miR-205 relative to U6B and miR-21 (Lebanony score), or only U6B (miR-205 ΔCqU6B) in 5 ng RNA for 49 AC and 28 SCC cases. For each type of measurement, A shows dot-plots of individual values along with histology-specific group means and standard deviations; B shows receiver operating characteristic curves, along the AUC with 95% confidence interval, for the miR-205 measurements to discriminate SCC from AC; and, C shows the measurement values in terms of molarity along with linear regression lines and their slopes (m) and Y intercepts (b) determined by the least squares fitting technique. Molarity equivalent of the Lebanony score could be determined for only 56 of the total 77 cases. SCC, squamous cell carcinoma; AC, adenocarcinoma; RT-PCR, reverse transcription-polymerase chain reaction; AUC, area under curve. Journal of Thoracic Oncology 2015 10, 446-453DOI: (10.1097/JTO.0000000000000423) Copyright © 2015 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 4 Expression of miR-205 and miR-375 in lung SCC and AC resectates. RT-PCR was used to quantify miR-205 relative to U6B and miR-21 (Lebanony score), and of miR-375 relative to U6B (miR-375 ΔCqU6B) in 5 ng RNA for 49 AC (gray) and 28 SCC (black) cases. Dotted lines indicate the median and SD values of miR-375 ΔCqU6B and Lebanony scores of the AC cases with scores greater than 2.5. SCC, squamous cell carcinoma; AC, adenocarcinoma; RT-PCR, RT-PCR, reverse transcription-polymerase chain reaction; SD, standard deviation. Journal of Thoracic Oncology 2015 10, 446-453DOI: (10.1097/JTO.0000000000000423) Copyright © 2015 International Association for the Study of Lung Cancer Terms and Conditions