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通过对合格胰液样本中人端粒酶逆转录酶信使核糖核酸的检测诊断胰腺腺癌

Diagnosis of pancreatic adenocarcinoma by detection of human telomerase reverse transcriptase messenger RNA in pancreatic juice with sample qualification.

作者信息

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

机构信息

Department of Molecular Genetics, Graduate School of Medical and Dental Sciences, Niigata University, Niigata 951-8122, Japan.

出版信息

Clin Cancer Res. 2001 Jul;7(7):1976-81.

Abstract

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, 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.

摘要

目的

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。

实验设计

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。

结果

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。

结论

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。

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