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Detection of Mutant K-ras in a Kindred With Hereditary Pancreatic Cancer by DGGE

David A. Crispin,* Ru Chen,* Michael B. Kimmey,** Teresa A. Brentnall**
* Departments of Pathology and
**Medicine, Division of Gastroenterology, University of Washington, Seattle, Washington.


Introduction

Pancreatic cancer is the fourth leading cause of cancer death in the U.S. and its frequency is rising. At the time of diagnosis, 9699% of patients are incurable and will shortly die. Current methods to evaluate patients for pancreatic adenocarcinoma include Endoscopic Retrograde Cholangiopancreatography (ERCP), abdominal CT, ultrasound, and serum markers. These methods of diagnosis are often insensitive or equivocal in early disease. Tumorigenesis is believed to involve the K-ras oncogene, DCC, p16, APC, bcl-2 and p53 tumor suppressor genes; screening for mutations or loss of heterozygosity in these genes may provide better diagnostic tests that are more sensitive and specific.1 The study of families in which cancer is inherited in an autosomal dominant fashion has provided considerable insight into understanding the molecular basis for pancreatic cancer. We have previously reported an extensive kindred in which pancreatic cancer is inherited in an autosomal dominant fashion and is associated with development of pancreatic insufficiency prior to the diagnosis of cancer.2 We have utilized the DCode system for denaturing gradient gel electrophoresis (DGGE) to identify patient samples with K-ras mutations.


Materials and Methods

Genomic DNA was isolated from tissue or from fluid obtained from patients at ERCP. K-ras exon 1 was amplified from genomic DNA using a thermal cycler
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