Thyroid Transcription Factor-1 Amplification and Expressions in Lung Adenocarcinoma Tissues and Pleural Effusions Predict Patient Survival and Prognosis

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Thyroid Transcription Factor-1 Amplification and Expressions in Lung Adenocarcinoma Tissues and Pleural Effusions Predict Patient Survival and Prognosis Xiao Li, MD, Liyan Wan, MD, Hong Shen, PhD, MD, Jian Geng, MD, Jing Nie, PhD, Guobao Wang, MD, Nan Jia, PhD, MD, Meng Dai, PhD, MD, Xiaoyan Bai, PhD, MD Journal of Thoracic Oncology Volume 7, Issue 1, Pages 76-84 (January 2012) DOI: 10.1097/JTO.0b013e318232b98a Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 Thyroid transcription factor-1 (TTF1) gene amplification in primary lung adenocarcinoma, lymph node metastasis, and malignant pleural effusions detected with nuclei microarray fluorescence in situ hybridization (NMA-FISH). A, Overview of a NMA slide (hematoxylin and eosin [H&E]). B, One NMA disk stained by diamidinophenyl-indole (DAPI, original magnification, ×200). C, TTF1 gene amplification in primary lung adenocarcinoma (63× oil). D, TTF1 gene amplification in lymph node metastasis (63× oil). E, TTF1 gene amplification in MPEs (63× oil). Red signals represent amplification of the NKX2-1 probe and green signals represent a chromosome 14 reference probe. Nuclei are stained in blue with DAPI. Journal of Thoracic Oncology 2012 7, 76-84DOI: (10.1097/JTO.0b013e318232b98a) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 Thyroid transcription factor-1 (TTF1) messenger RNA (mRNA) expression level was twofold higher in lung adenocarcinomas with TTF1 gene amplification. A, Quantitative real-time polymerase chain reaction (PCR) showed increased TTF1 mRNA expression level in lung adenocarcinomas with TTF1 gene amplification compared with normal lung and lung adenocarcinomas without TTF1 gene amplification. B, Representative image of mRNA expression level of seven snap-frozen tumor tissues and matched normal lung tissues detected by reverse transcriptase (RT)-PCR. T4, T5, and T6 represented three lung adenocarcinoma tissues with TTF1 gene amplification. Journal of Thoracic Oncology 2012 7, 76-84DOI: (10.1097/JTO.0b013e318232b98a) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 Thyroid transcription factor-1 (TTF1) protein and messenger RNA (mRNA) expressions were positive in papillary subtype of primary lung adenocarcinomas and lymph node metastases and negative in invasive mucinous primary lung adenocarcinomas (scale bar: 100μm). A, Positive TTF1 protein expression in papillary lung adenocarcinoma. B, Positive TTF1 mRNA expression in papillary lung adenocarcinoma. C, Negative TTF1 protein expression in invasive mucinous lung adenocarcinoma. D, Negative TTF1 mRNA expression in invasive mucinous lung adenocarcinoma. E, Positive TTF1 protein expression in lymph node metastasis. F, Positive TTF1 mRNA expression in lymph node metastasis. Journal of Thoracic Oncology 2012 7, 76-84DOI: (10.1097/JTO.0b013e318232b98a) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 4 Representative images of thyroid transcription factor-1 (TTF1) protein and messenger RNA (mRNA) expressions in the MPE of patients with lung cancer (scale bar: 100 μm). TTF1 expressions were positive in five cases of MPE (three invasive mucinous types and two papillary types). A-1, Hematoxylin and eosin (H&E); A-2, negative TTF1 protein; and A-3, mRNA expressions in papillary lung adenocarcinoma. B-1, H&E; B-2, positive TTF1 protein; and B-3, mRNA expressions in invasive mucinous lung adenocarcinoma. C, Representative TTF1 expressions in MPE detected by Western blot. Journal of Thoracic Oncology 2012 7, 76-84DOI: (10.1097/JTO.0b013e318232b98a) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 5 Kaplan-Meier estimates of overall survival among patients with lung adenocarcinomas according to thyroid transcription factor-1 (TTF1) expressions and TTF1 amplification. A, There was a trend toward a worse overall survival for patients with lung adenocarcinoma with positive TTF1 expressions and TTF1 gene amplification compared with patients with TTF1 expressions and no TTF1 amplification (median: 48.4 versus 74.3 months, p = 0.068). B, The overall survival time was longer for patients with lung adenocarcinoma with positive TTF1 expressions, than for patients with negative TTF1 expressions (median: 72.6 versus 42.5 months, p < 0.001). C, The overall survival time was significantly longer for patients with positive TTF1 expressions in MPE compared with patients with negative TTF1 expressions (median: 72.6 versus 43.9 months, p = 0.045). Journal of Thoracic Oncology 2012 7, 76-84DOI: (10.1097/JTO.0b013e318232b98a) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions