Aberrant activation of the Wnt signaling pathway has been reported during neoplastic progression in Barrett's esophagus (BE). However, mutations in APC and CTNNB1 genes were rarely observed. In this study, expression pattern of Wnt ligands, Frizzled receptors and APC, as well as the methylation status of the APC, SFRP1 and SFRP2 promoter genes were investigated in normal esophageal mucosa and in preneoplastic and neoplastic lesions of BE patients. Promoter methylation of APC was found in all BE samples and in 95% of esophageal adenocarcinomas (EAC). Full methylation of APC correlated with lack of expression. In EAC, nuclear translocation of b-catenin was observed regardless of the expression of APC. WNT2 expression was higher in dysplasia and EAC than in BE, with 20/26 (77%) of the EAC showing high expression of WNT2. SFRP1 methylation occurred in all BE samples and in 96% of EAC, while SFRP2 was methylated in 73% of the normal squamous esophageal mucosa samples. In conclusion, (1) alterations of key regulators of the Wnt signaling are frequent in the pathogenesis of BE; (2) the APC and SFRP1 genes are inactivated by promoter methylation in BE; (3) the WNT2 gene is upregulated along the progression from low-grade dysplasia to EAC.

Barrett's associated oesophageal adenocarcinoma (EAC) is one of the most rapidly increasing malignancies in Western countries. Because of its poor prognosis, management of this disease through screening of Barrett's oesophagus (BE) patients and identification of those with a high risk of developing an adenocarcinoma seems a promising approach. Early molecular markers of malignant transformation might contribute to such screening approaches. Gene promoter methylation analysis was performed on normal, pre-neoplastic, and neoplastic lesions from BE patients. All lesions of interest were sampled by microdissection from formalin-fixed paraffin-embedded tissue sections. We found that, in 27 adenocarcinomas, APC, TIMP3, TERT, CDKN2A, and SFRP1 promoters were methylated in 93%, 65%, 64%, 48%, and 91%, respectively; in contrast MLH1, RASSF1, RARB, CDH1, and FHIT promoters were methylated in less than 5% of the tumours. In BE mucosa from patients who had progressed to adenocarcinoma (12 samples), APC, TIMP3, and TERT promoters were hypermethylated in 100%, 91%, and 92% of cases, whereas in BE mucosa from patients who had not progressed (16 samples) methylation was found only in 36%, 23%, and 17%, respectively. Furthermore, the epigenetic profile of BE with and without EAC differed significantly with, respectively, 81% and 26% of the PCR samples showing promoter hypermethylation for APC, TIMP3, and TERT (p < 0.0001). Promoter methylation of CDKN2A was infrequently detected in BE samples, while SFRP1 methylation was observed in all samples. Our results suggest that promoter methylation profiling of BE using multiple target genes including APC, TIMP3, and TERT might be used as a predictive marker for increased EAC risk.

Extrathoracic metastases of thymic neoplasms are very rare 1,2 (incidence is believed to be around 3-6%), and their cytologic features have seldom been reported. 1-5 The most common subtype associated with metastasis is thymic carcinoma, followed by type B thymoma. 1,2 The recognition of thymic metastasis is challenging because of its rarity and because of its potency to mimic tumors from other sites. We report the case of a metastatic type B thymoma in supraclavicular and mediastinal lymph nodes in a 69-year-old male diagnosed on fine-needle aspiration biopsy (FNAB). The patient had a previous history of type B (mixed or lympho-epithelial) thymoma with myasthenia treated with corticosteroids, surgery, and radiotherapy 12 years earlier. The primary tumor, partially calcified, measured 7 3 6.5 3 3 cm and involved the pericardium and periaortic fat. Surgical margins were free of tumor. The patient developed left diaphragm and left vocal cord palsy after surgery. In 2012, he was admitted to the hospital because of recurrent bilateral bronchopneumonia due to multiresistant Pseudomonas aeruginosa. The investigations revealed mediastinal (upper and middle) and left supraclavicular lymphadenopathies with left carotid artery involvement. FNAB of lymphadenopathies from both localizations was performed and liquidbased preparations (Thinprep V R ) were made. The aspirates were cellular, composed of cohesive epithelioid to spindle