巴雷特食管的非内镜筛查生物标志物：从微阵列分析到临床应用

Non-endoscopic screening biomarkers for Barrett's oesophagus: from microarray analysis to the clinic.

作者信息

Lao-Sirieix P, Boussioutas A, Kadri S R, O'Donovan M, Debiram I, Das M, Harihar L, Fitzgerald R C

机构信息

MRC-Cancer Cell Unit, Hutchison-MRC Research Centre, Cambridge CB22 0XZ, UK.

出版信息

Gut. 2009 Nov;58(11):1451-9. doi: 10.1136/gut.2009.180281. Epub 2009 Aug 2.

DOI:10.1136/gut.2009.180281

PMID:19651633

Abstract

BACKGROUND AND AIMS

Barrett's oesophagus predisposes to oesophageal adenocarcinoma but the majority of patients are undiagnosed. A novel non-endoscopic cytological screening device, called a capsule sponge, makes population-based screening for the disease a feasible option. However, due to the mixed cell population retrieved by the capsule sponge, biomarkers specific for Barrett's oesophagus are required.

METHODS

Three publically available microarray datasets were used to identify putative biomarkers present in Barrett's oesophagus but absent from normal oesophagus and gastric mucosa. Validation was performed by qPCR (n = 10 each of normal oesophagus, Barrett's oesophagus, gastric mucosa) and immunohistochemistry (normal oesophagus, n = 20; Barrett's oesophagus, n = 21; gastric mucosa, n = 24; duodenum, n = 18). The biomarker was then prospectively evaluated on capsule sponge specimens from 47 patients with Barrett's oesophagus and 99 healthy controls.

RESULTS

2/14 genes identified, dopa decarboxylase (DDC) and Trefoil factor 3 (TFF3), were confirmed by qPCR to be upregulated in Barrett's oesophagus compared to normal oesophagus (p<0.01) and gastric mucosa (p<0.01 and p<0.05, respectively). Immunohistochemistry confirmed that DDC protein expression was restricted to Barrett's oesophagus but was confined to <1% of the cells within the crypt compartment. TFF3 protein was expressed to high levels at the luminal surface of Barrett's oesophagus compared to absent expression in normal oesophagus and gastric mucosa (p<0.001). Using the capsule sponge 36/46 patients with Barrett's oesophagus (one inadequate sample) and 6/96 controls were positive for TFF3 giving a sensitivity of 78% and a specificity of 94%.

CONCLUSIONS

TFF3 is a promising marker for Barrett's oesophagus screening since it is expressed at the luminal surface of Barrett's oesophagus but not in adjacent tissue types and may be applied to a non-endoscopic screening device.

摘要

背景与目的

巴雷特食管易引发食管腺癌，但大多数患者未被诊断出来。一种名为胶囊海绵的新型非内镜细胞学筛查设备，使基于人群的该疾病筛查成为一种可行的选择。然而，由于胶囊海绵获取的细胞群体混合，需要巴雷特食管特异性的生物标志物。

方法

使用三个公开可用的微阵列数据集来识别存在于巴雷特食管但在正常食管和胃黏膜中不存在的假定生物标志物。通过qPCR（正常食管、巴雷特食管、胃黏膜各10例）和免疫组织化学（正常食管20例；巴雷特食管21例；胃黏膜24例；十二指肠18例）进行验证。然后对47例巴雷特食管患者和99例健康对照的胶囊海绵标本进行该生物标志物的前瞻性评估。

结果

通过qPCR确认，所鉴定的14个基因中的2个，即多巴脱羧酶（DDC）和三叶因子3（TFF3），在巴雷特食管中相对于正常食管（p<0.01）和胃黏膜（分别为p<0.01和p<0.05）上调。免疫组织化学证实，DDC蛋白表达仅限于巴雷特食管，但局限于隐窝区不到1%的细胞内。与正常食管和胃黏膜中无表达相比，TFF3蛋白在巴雷特食管的腔面高水平表达（p<0.001）。使用胶囊海绵，46例巴雷特食管患者中的36例（1例样本不足）和96例对照中的6例TFF3呈阳性，敏感性为78%，特异性为94%。