PMC4512946_01
Epidermal growth factor receptor (EGFR) T790M Mutation in Two Patients with Lung Cancer. Panel A, shows the pedigree of the patient with the germ-line EGFR T790M mutation (the patient is indicated by the arrow, and lung cancer is indicated by solid symbols). Panel B, shows a diagram of the patient's lungs illustrating the three synchronous lung tumors and representative microphotographs from histologic analysis (hematoxylin and eosin, magnification X400). Tumor 1 (T1) was an invasive adenocarcinoma located in the right lower lobe, tumor 2 (T2) was a pure (noninvasive) bronchioloalveolar carcinoma (BAC) located in the right middle lobe, and tumor 3 (T3) was a large-cell neuroendocrine carcinoma (LCNEC) located in the left lower lobe with hilar lymph node metastasis. Deoxyribonucleic acid (DNA) was extracted from about 1000 microdissected cells from four sites in T1 and 1 site each from the T2 and T3 lymph node metastases (red circles) and then examined for EGFR mutations in exons 18 to 21. In addition, microdissected connective pleural tissue (two sites), nonmetastatic lymph node (two sites), and normal bronchial epithelium adjacent to T1 and T2 (7 sites) were also examined (sites not indicated). All the specimens, nonmalignant and malignant, demonstrated the T790M mutation. Panel C, shows examples of nucleotide sequence tracings illustrating heterozygosity with respect to the T790M mutation in DNA extracted from the peripheral mononuclear blood cells and the tissue sites examined. Panel D, shows the pedigree of the patient with the somatic EGFR T790M mutation (the patient is indicated by the arrow, and lung cancer is indicated by the solid symbols). Panel E, shows a diagram of the patient's lungs illustrating the tumor and representative microphotographs from histologic analysis of the normal epithelium and tumor (hematoxylin and eosin, magnification X400). In this case, the T790M mutation was detected in four of the four tumor sites (T, red circles) and three of the five normal bronchial epithelium sites (NE, mutant: red circles, wild-type: yellow circles). Microdissected tissue from two pleural (P, green circles) and lymph node (LN, blue circles) sites without tumor involvement had the wild-type allele. Panel F, shows examples of nucleotide sequence tracings illustrating heterozygosity with respect to the T790M variant in normal bronchiolar epithelium and tumor specimen sites. The lymph node sites and pleural tissues had the wild-type allele.
nihms-707884-f0001.jpg
PMC4512946_02_nihms-707884-f0001.jpg
F1
T790M-associated lung cancer occurred in a 62-year-old female never smoker diagnosed with stage IA adenocarcinoma; no other EGFR activating mutation was seen (Figures 1D-F). The T790M mutation was not detected in the DNA extracted from multiple microdissected tissue samples, including nonmalignant pleural connective tissue, lung hilar lymph nodes, and breast connective tissue obtained during a previous breast cancer resection showing that mutation was somatic.
PMC4512946_02
Epidermal growth factor receptor (EGFR) T790M Mutation in Two Patients with Lung Cancer. Panel A, shows the pedigree of the patient with the germ-line EGFR T790M mutation (the patient is indicated by the arrow, and lung cancer is indicated by solid symbols). Panel B, shows a diagram of the patient's lungs illustrating the three synchronous lung tumors and representative microphotographs from histologic analysis (hematoxylin and eosin, magnification X400). Tumor 1 (T1) was an invasive adenocarcinoma located in the right lower lobe, tumor 2 (T2) was a pure (noninvasive) bronchioloalveolar carcinoma (BAC) located in the right middle lobe, and tumor 3 (T3) was a large-cell neuroendocrine carcinoma (LCNEC) located in the left lower lobe with hilar lymph node metastasis. Deoxyribonucleic acid (DNA) was extracted from about 1000 microdissected cells from four sites in T1 and 1 site each from the T2 and T3 lymph node metastases (red circles) and then examined for EGFR mutations in exons 18 to 21. In addition, microdissected connective pleural tissue (two sites), nonmetastatic lymph node (two sites), and normal bronchial epithelium adjacent to T1 and T2 (7 sites) were also examined (sites not indicated). All the specimens, nonmalignant and malignant, demonstrated the T790M mutation. Panel C, shows examples of nucleotide sequence tracings illustrating heterozygosity with respect to the T790M mutation in DNA extracted from the peripheral mononuclear blood cells and the tissue sites examined. Panel D, shows the pedigree of the patient with the somatic EGFR T790M mutation (the patient is indicated by the arrow, and lung cancer is indicated by the solid symbols). Panel E, shows a diagram of the patient's lungs illustrating the tumor and representative microphotographs from histologic analysis of the normal epithelium and tumor (hematoxylin and eosin, magnification X400). In this case, the T790M mutation was detected in four of the four tumor sites (T, red circles) and three of the five normal bronchial epithelium sites (NE, mutant: red circles, wild-type: yellow circles). Microdissected tissue from two pleural (P, green circles) and lymph node (LN, blue circles) sites without tumor involvement had the wild-type allele. Panel F, shows examples of nucleotide sequence tracings illustrating heterozygosity with respect to the T790M variant in normal bronchiolar epithelium and tumor specimen sites. The lymph node sites and pleural tissues had the wild-type allele.
nihms-707884-f0001.jpg
PMC4512946_02_nihms-707884-f0001.jpg
F1
T790M-associated lung cancer occurred in a 62-year-old female never smoker diagnosed with stage IA adenocarcinoma; no other EGFR activating mutation was seen (Figures 1D-F). The T790M mutation was not detected in the DNA extracted from multiple microdissected tissue samples, including nonmalignant pleural connective tissue, lung hilar lymph nodes, and breast connective tissue obtained during a previous breast cancer resection showing that mutation was somatic.