直接检测血清游离 BMI-1 mRNA 及其对结直肠癌的潜在诊断和预后价值。

Direct serum assay for cell-free bmi-1 mRNA and its potential diagnostic and prognostic value for colorectal cancer.

机构信息

Department of Clinical Laboratory, Qilu Hospital, Shandong University, Jinan, Shandong Province, China.

Department of Gastroenterology, Qilu Hospital, Shandong University, Jinan, Shandong Province, China.

出版信息

Clin Cancer Res. 2015 Mar 1;21(5):1225-33. doi: 10.1158/1078-0432.CCR-14-1761. Epub 2014 Dec 29.

DOI:10.1158/1078-0432.CCR-14-1761

PMID:25547677

Abstract

PURPOSE

Cell-free Bmi-1 mRNA is stably detectable in the serum/plasma and is associated with the development and progression of some tumors. Previous methods detecting extracellular Bmi-1 mRNA with RNA extraction are inefficient. This study developed a novel reverse transcription quantitative PCR (RT-qPCR) approach directly applied in serum (RT-qPCR-D) to quantify Bmi-1 mRNA, and assessed its diagnostic and prognostic potential in colorectal cancer.

EXPERIMENTAL DESIGN

The feasibility of the RT-qPCR-D method was first analyzed in 50 serum samples. Then, using the RT-qPCR-D method, Bmi-1 mRNA expression was validated in serum from an independent cohort of patients with 87 normal colonoscopy, 76 hyperplastic polyp, 82 inflammatory bowel disease, 68 adenoma, and 158 colorectal cancer. Receiver operating characteristic (ROC) curves and Cox analyses were used to evaluate its diagnosis and prognosis value, respectively.

RESULTS

In a pilot study, levels of Bmi-1 mRNA were increased in colorectal cancer serum samples detected by RT-qPCR-D and significantly associated with results obtained by RT-qPCR. In a validation cohort, serum Bmi-1 mRNA levels were significantly elevated in the colorectal cancer group and the adenoma group when compared with other groups. The area under ROC curve distinguishing colorectal cancer from benign colorectal diseases was 0.888, with 72.2% sensitivity and 94.9% specificity, which was superior to carcinoembryogenic antigen. Bmi-1 mRNA levels were significantly associated with survival. Cox analysis indicated Bmi-1 mRNA was an independent prognostic factor for overall survival.

CONCLUSIONS

Detection of cell-free Bmi-1 mRNA in serum by RT-qPCR-D is a simple and noninvasive approach and may be used for colorectal cancer diagnosis and prognosis.

摘要

目的

无细胞 Bmi-1 mRNA 在血清/血浆中稳定可检测，并与一些肿瘤的发生和发展相关。之前使用 RNA 提取检测细胞外 Bmi-1 mRNA 的方法效率不高。本研究开发了一种新的逆转录定量 PCR（RT-qPCR）方法，直接应用于血清（RT-qPCR-D）来定量 Bmi-1 mRNA，并评估其在结直肠癌中的诊断和预后潜力。

实验设计

首先在 50 份血清样本中分析 RT-qPCR-D 方法的可行性。然后，使用 RT-qPCR-D 方法，在来自 87 名正常结肠镜检查、76 名增生性息肉、82 名炎症性肠病、68 名腺瘤和 158 名结直肠癌患者的独立队列血清中验证 Bmi-1 mRNA 的表达。使用受试者工作特征（ROC）曲线和 Cox 分析分别评估其诊断和预后价值。

结果

在一项初步研究中，通过 RT-qPCR-D 检测到结直肠癌血清样本中的 Bmi-1 mRNA 水平升高，且与 RT-qPCR 结果显著相关。在验证队列中，与其他组相比，结直肠癌组和腺瘤组的血清 Bmi-1 mRNA 水平显著升高。区分结直肠癌与良性结直肠疾病的 ROC 曲线下面积为 0.888，灵敏度为 72.2%，特异性为 94.9%，优于癌胚抗原。Bmi-1 mRNA 水平与生存显著相关。Cox 分析表明 Bmi-1 mRNA 是总生存期的独立预后因素。