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Diagnosis of Pancreatic Adenocarcinoma by Detection of Human Telomerase Reverse Transcriptase Messenger RNA in Pancreatic Juice with Sample Qualification¹

Department of Molecular Genetics [K. S., T. S., Y. A., S. S., M. N., H. M., H. K., Y. M., N. W., T. K., M. I., H. A.], Department of Regenerative and Transplant Medicine [Y. S.], Graduate School of Medical and Dental Sciences, Niigata University, Niigata 951-8122, and The Division of Internal Medicine, Shibata Prefecture Hospital, Shibata 957-0052 [T. S.], Japan

Purpose: We evaluated the diagnostic efficacy of detection of human telomerase reverse transcriptase (hTERT) message, a catalytic domain of human telomerase, in endoscopic retrograde pancreatography (ERP)-derived pancreatic juice.

Experimental Design: Both hTERT and CD25 expression were detected by reverse transcription-PCR (RT-PCR) in 17 patients with pancreatic adenocarcinoma (PC), 12 patients with chronic pancreatitis (CP), and 7 patients with no ERP abnormality (N). In the same patients, ß-actin message was semiquantified by competitive RT-PCR. K-ras codon 12 mutations were concomitantly analyzed by enriched PCR-SSCP in 11 and 7 PC and CP cases, respectively.

Results: Expression of hTERT was detected in 88% of PC cases and 17% of CP cases but not in the normal control (N). Alterations in K-ras were detected in 73% of PC cases and 57% of CP cases, respectively. ß-Actin mRNA was expressed in >3.0 x 10¹ copies/µl in all but two PC cases in which hTERT mRNA was not detected. CD25-positive and -negative peripheral lymphocytes were isolated from a normal volunteer using a fluorescent activating cell sorter. The hTERT message was detected in CD25-positive peripheral lymphocytes and in 18, 25, and 0% of the pancreatic juice samples from PC, CP, and N cases, respectively. All CP cases expressing hTERT message were also CD25 positive.

Conclusions: These results suggest that detection of hTERT mRNA in pancreatic juice is a powerful tool to discriminate PC from CP, particularly when the samples are qualified against ß-actin mRNA levels and contaminating CD25-positive lymphocytes.

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