Seki K, Suda T, Aoyagi Y, Sugawara S, Natsui M, Motoyama H, Shirai Y, Sekine T, Kawai H, Mita Y, Waguri N, Kuroiwa T, Igarashi M, Asakura H
Department of Molecular Genetics, Graduate School of Medical and Dental Sciences, Niigata University, Niigata 951-8122, Japan.
Clin Cancer Res. 2001 Jul;7(7):1976-81.
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.
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.
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.
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.
我们评估了在内镜逆行胰胆管造影(ERP)获取的胰液中检测人端粒酶逆转录酶(hTERT)信息(人端粒酶的催化结构域)的诊断效能。
通过逆转录聚合酶链反应(RT-PCR)检测了17例胰腺腺癌(PC)患者、12例慢性胰腺炎(CP)患者和7例ERP无异常(N)患者的hTERT和CD25表达。在相同患者中,通过竞争性RT-PCR对β-肌动蛋白信息进行半定量分析。分别对11例PC病例和7例CP病例通过富集PCR-SSCP同时分析K-ras密码子12突变情况。
88%的PC病例和17%的CP病例检测到hTERT表达,但正常对照组(N)未检测到。PC病例和CP病例中分别有73%和57%检测到K-ras改变。除两例未检测到hTERT mRNA的PC病例外,所有病例的β-肌动蛋白mRNA表达均>3.0×10¹拷贝/微升。使用荧光激活细胞分选仪从一名正常志愿者中分离出CD25阳性和阴性外周淋巴细胞。在CD25阳性外周淋巴细胞以及分别来自PC、CP和N病例的18%、25%和0%的胰液样本中检测到hTERT信息。所有表达hTERT信息的CP病例也为CD25阳性。
这些结果表明,检测胰液中的hTERT mRNA是区分PC和CP的有力工具,特别是当样本根据β-肌动蛋白mRNA水平和污染的CD25阳性淋巴细胞进行校正时。