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Identification of K-ras Mutations in Pancreatic Juice in the Early Diagnosis of Pancreatic Cancer

Philippe Berthelemy; Michele Bouisson; Jean Escourrou; Nicole Vaysse; Jean Louis Rumeau; and Lucien Pradayrol
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From Institut National de la Sante et de la Recherche Medicale, Toulouse, France. Request for Reprints: Lucien Pradayrol, PhD, Unite 151, Institut National de la Sante et de la Recherche Medicale, Biologie et Pathologie Digestive, Bt L3, CHU. Rangueil, Institut Louis Bugnard, 1, Avenue J. Poulhes, F31054 Toulouse CEDEX, France. Acknowledgments: The authors thank Eli Yardeni. Grant Support: In part by research grants from the Delegation a la Recherche Clinique. Dr. Berthelemy is supported by Bourse de Recherche du Club Francais du Pancreas (Laboratoires Eurorga-Jouveinal).

Copyright ©2004 by the American College of Physicians

Ann Intern Med. 1995;123(3):188-191. doi:10.7326/0003-4819-123-3-199508010-00005
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Objective: To develop an early diagnostic test for pancreatic cancer based on the identification of K-ras mutations in pure pancreatic juice collected during endoscopic retrograde pancreatography.

Design: Prospective study with masked comparison. The standard criteria for the diagnosis of pancreatic cancer were pancreatography or surgery (or both) and histopathology, with follow-up ranging from 6 to 40 months.

Setting: Referral center.

Patients: 24 patients with no pancreatic disease (group 1); 29 patients with nontumoral pancreatic disease (group 2); and 22 patients with pancreatic tumor (group 3). Endoscopic ductal aspiration of cells or brush cytology was done on patients having endoscopic retrograde pancreatography for diagnostic or therapeutic reasons.

Main Outcome Measure: Confirmation of mutation rates in patients with pancreatic cancer.

Results: K-ras gene analysis was done by polymerase chain reaction-mediated restriction fragment length polymorphism analysis and direct sequencing. All patients from groups 1 and 2 (n = 53) had a normal sequence for the K-ras 12th codon (group 1, 0% [95% CI, 0% to 14%]; group 2, 0% [CI, 0% to 12%]). Mutations were seen in 17 of the 22 patients in group 3 (77% [CI, 55% to 92%]). Two of the 17 had no evidence of pancreatic cancer when K-ras was first studied. One had chronic abdominal pain and the other presented with acute pancreatitis. Both were initially free of any pancreatic mass, but they developed tumors 18 and 40 months, respectively, after the K-ras mutations were identified.

Conclusion: Identification of K-ras mutations in samples of pancreatic juice may be useful in differentiating between pancreatic cancer and noncancerous pancreatic diseases. K-ras mutation can precede clinical evidence of pancreatic cancer, but the clinical implications of this finding need further study.


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Figure 1.
Detection of K-ras 12th codon mutation in pancreatic juice by polymerase chain reaction-mediated restriction length fragment polymorphism analysis.

Polymerase chain reaction amplification produced a 165-bp fragment that contained a BstN1 cleavage site. This site was absent in mutant amplified K-ras fragment. Arrows indicate the positions of the nondigested (165 bp) and the digested wild-type (136 bp) fragment. (1, 1′,2, 2′) = normal K-ras gene fragment; (3, 3′) = DNA sample showing both the mutant (165 bp) and the wild-type (136 bp) fragments after BstN1 digestion.

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