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Evaluation of clinical relevance of examining K-ras, p16 and p53 mutations along with allelic losses at 9p and 18q in EUS-guided fine needle aspiration samples of patients with chronic pancreatitis and pancreatic cancer

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World Journal of Gastroenterology 2007年第27期13卷 3714-3720页

作者： C Salek l benesova M Zavoral V Nosek l Kasperova M Ryska R Strnad E Traboulsi M Minarik Department of Gastroenterology Central Military Hospital and 1~(st) Medical Faculty of Charles University Prague Czech Republic Laboratory for Molecular Genetics and Oncology Genomac International Ltd. Prague Czech Republic Department of Gastroenterology Institute for Postgraduate Medical Education Prague Czech Republic laboratory for Molecular Genetics and Oncology Genomac International Ltd. Prague Czech Republic Department of Gastroenterology Central Military Hospital and 1~(st) Medical Faculty of Charles University Prague Czech Republic Department of Gastroenterology Institute for Postgraduate Medical Education Prague Czech Republic Department of Gastroenterology Central Military Hospital and 1~(st) Medical Faculty of Charles University Prague Czech Republic Department of Surgery Central Military Hospital and 2nd Medical Faculty of Charles University Prague Czech Republic Department of Pathology Central Military Hospital Prague Czech Republic

AIM： To establish an optimum combination of molecular markers resulting in best overall diagnostic sensitivity and specificity for evaluation of suspicious pancreatic mass. METHODS： Endoscopic ultrasound （EUS）-guided fine needle aspiration cytology （FNA） was performed on 101 consecutive patients （63 males, 38 females, 60 ± 12 years; 81 with subsequently diagnosed pancreatic cancer, 20 with chronic pancreatitis） with focal pancreatic mass. Samples were evaluated on-site by an experienced cytopathologist. DNA was extracted from Giemsa stained cells selected by laser microdissection and the presence of K-ras, p53 and p16 somatic mutations was tested by cycling-gradient capillary electrophoresis （CGCE） and single-strand conformation polymorphism （SSCP） techniques. In addition, allelic losses of tumor suppressor genes p16 （INK4, CDKN2A） and DPC4 （MADH4, SMAD4） were detected by monitoring the loss of heterozygosity （lOH） at 9p and 18q, respectively. RESUlTS： Sensitivity and specificity of EUS-guided FNA were 75% and 85%, positive and negative predictive value reached 100%. The remaining 26% samples were assigned as inconclusive. Testing of molecular markers revealed sensitivity and specificity of 70% and 100% for K-ras mutations （P 〈 0.001）, 24% and 90% for p53 mutations （NS）, 13% and 100% for p16 mutations （NS）, 85% and 64% for aUelic losses at 9p （P 〈 0.001） and 78% and 57% for allelic losses at 18q （P 〈 0.05）. When tests for different molecular markers were combined, the best results were obtained with K-ras ＋ lOH at 9p （92% and 64%, P 〈 0.001）, K-ras ＋ lOH at 18q （92% and 57%, P 〈 0.001）, and K-ras ＋ lOH 9q ＋ lOH 18q （96% and 43%, P 〈 0.001）. When the molecular markers were used as complements to FNA cytology to evaluate inconclusive samples only, the overall sensitivity of cancer detection was 100% in all patients enrolled in the study. CONClUSION： EUS-guided FNA cytology combined with screening of K-ras mutations and allelic losses of tumor suppressors p16 and

关键词： Pancreatic cancer Chronic pancreatitis Endoscopic ultrasound-guided fine-needle aspiration Molecular markers loss of heterozygosity

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