title: Diagnosis of pancreatic adenocarcinoma by detection of human telomerase reverse transcriptase messenger RNA in pancreatic juice with sample qualification.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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, beta-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. beta-Actin mRNA was expressed in >3.0 x 10(1) copies/microl 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 beta-actin mRNA levels and contaminating CD25-positive lymphocytes.
authors.affiliation: Department of Molecular Genetics, Graduate School of Medical and Dental Sciences, Niigata University, Niigata 951-8122, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Seki
authors.forename: T
authors.initials: T
authors.lastname: Suda
authors.forename: Y
authors.initials: Y
authors.lastname: Aoyagi
authors.forename: S
authors.initials: S
authors.lastname: Sugawara
authors.forename: M
authors.initials: M
authors.lastname: Natsui
authors.forename: H
authors.initials: H
authors.lastname: Motoyama
authors.forename: Y
authors.initials: Y
authors.lastname: Shirai
authors.forename: T
authors.initials: T
authors.lastname: Sekine
authors.forename: H
authors.initials: H
authors.lastname: Kawai
authors.forename: Y
authors.initials: Y
authors.lastname: Mita
authors.forename: N
authors.initials: N
authors.lastname: Waguri
authors.forename: T
authors.initials: T
authors.lastname: Kuroiwa
authors.forename: M
authors.initials: M
authors.lastname: Igarashi
authors.forename: H
authors.initials: H
authors.lastname: Asakura
mesh_terms: D000199:Actins / Q000235:genetics; D000230:Adenocarcinoma / Q000175:diagnosis* / Q000235:genetics; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D003062:Codon / Q000235:genetics; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000737:chemistry / Q000235:genetics; D004268:DNA-Binding Proteins; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D011905:Genes, ras / Q000235:genetics; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D010189:Pancreatic Juice / Q000201:enzymology* / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000175:diagnosis* / Q000235:genetics; D010195:Pancreatitis / Q000175:diagnosis / Q000235:genetics; D018807:Polymorphism, Single-Stranded Conformational; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D015375:Receptors, Interleukin-2 / Q000235:genetics; D019098:Telomerase / Q000235:genetics*
chemical_list: D000199:Actins; D003062:Codon; D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D015375:Receptors, Interleukin-2; D019098:Telomerase