新型基于血液的 microRNA 生物标志物panel 用于胰腺癌的早期诊断。
Novel blood-based microRNA biomarker panel for early diagnosis of pancreatic cancer.
机构信息
Ganepola AP Ganepola, John R Rutledge, Paritosh Suman, David H Chang, Center for Cancer Research and Genomic Medicine, The Valley Hospital, Paramus, NJ 07652, United States.
出版信息
World J Gastrointest Oncol. 2014 Jan 15;6(1):22-33. doi: 10.4251/wjgo.v6.i1.22.
AIM
To develop a panel of blood-based diagnostic biomarkers consisting of circulating microRNAs for the detection of pancreatic cancer at an early stage.
METHODS
Blood-based circulating microRNAs were profiled by high throughput screening using microarray analysis, comparing differential expression between early stage pancreatic cancer patients (n = 8) and healthy controls (n = 11). A panel of candidate microRNAs was generated based on the microarray signature profiling, including unsupervised clustering and statistical analysis of differential expression levels, and findings from the published literature. The selected candidate microRNAs were then confirmed using TaqMan real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) to further narrow down to a three-microRNA diagnostic panel. The three-microRNA diagnostic panel was validated with independent experimental procedures and instrumentation of RT-qPCR at an independent venue with a new cohort of cancer patients (n = 11), healthy controls (n = 11), and a group of high risk controls (n = 11). Receiver operating characteristic curve analysis was performed to assess the diagnostic capability of the three-microRNA panel.
RESULTS
In the initial high throughput screening, 1220 known human microRNAs were screened for differential expression in pancreatic cancer patients versus controls. A subset of 42 microRNAs was then generated based on this data analysis and current published literature. Eight microRNAs were selected from the list of 42 targets for confirmation study, and three-microRNAs, miR-642b, miR-885-5p, and miR-22, were confirmed to show consistent expression between microarray and RT-qPCR. These three microRNAs were then validated and evaluated as a diagnostic panel with a new cohort of patients and controls and found to yield high sensitivity (91%) and specificity (91%) with an area under the curve of 0.97 (P < 0.001). Compared to the CA19-9 marker at 73%, the three-microRNA panel has higher sensitivity although CA19-9 has higher specificity of 100%.
CONCLUSION
The identified panel of three microRNA biomarkers can potentially be used as a diagnostic tool for early stage pancreatic cancer.
目的
开发一种基于血液的诊断生物标志物panel，由循环 microRNAs 组成，用于早期检测胰腺癌。
方法
使用微阵列分析进行高通量筛选，比较早期胰腺癌患者（n=8）与健康对照者（n=11）之间的差异表达，以分析基于血液的循环 microRNAs。基于微阵列特征谱分析，包括无监督聚类和差异表达水平的统计分析，并结合文献中的发现，生成候选 microRNAs panel。使用 TaqMan 实时定量逆转录聚合酶链反应（RT-qPCR）验证所选候选 microRNAs，进一步缩小至三 microRNA 诊断 panel。使用独立实验程序和仪器在独立场所对三 microRNA 诊断 panel 进行验证，使用新的癌症患者队列（n=11）、健康对照者（n=11）和高危对照组（n=11）进行 RT-qPCR。进行接收者操作特征曲线分析以评估三 microRNA 面板的诊断能力。
结果
在最初的高通量筛选中，筛选了 1220 个已知的人类 microRNAs，以检测胰腺癌患者与对照组之间的差异表达。然后基于该数据分析和当前发表的文献，生成了一个 42 个 microRNAs 的子集。从 42 个靶标列表中选择了 8 个 microRNAs 进行确认研究，miR-642b、miR-885-5p 和 miR-22 这三个 microRNAs 被证实与微阵列和 RT-qPCR 之间的表达一致。然后使用新的患者和对照者队列对这三个 microRNAs 进行验证和评估，作为诊断 panel，发现具有较高的灵敏度（91%）和特异性（91%），曲线下面积为 0.97（P<0.001）。与 CA19-9 标志物的 73%相比，虽然 CA19-9 的特异性为 100%，但三 microRNA 面板具有更高的灵敏度。
结论
所鉴定的三 microRNA 生物标志物 panel 可潜在用作早期胰腺癌的诊断工具。
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