Salivary transcriptome diagnostics for oral cancer detection
Salivary transcriptome diagnostics for oral cancer detection
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DOI：
10.1158/1078-0432.ccr-04-1167
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发表时间：
2004-12-15
影响因子：
11.5
通讯作者：
Wong, DT
中科院分区：
文献类型：
--
作者：
Li, Y;St John, MAR;Wong, DT
目的：口服液(唾液)满足了对非侵入性、可获得性和高效诊断介质的需求。最近发现，唾液中可以可靠地检测到大量人类RNA，这引发了一种新的临床方法，唾液转录组诊断。本研究的目的是以口腔鳞状细胞癌(OSCC)为基础疾病来评价这一新方法的诊断价值。实验设计：收集来自年龄、性别和吸烟史匹配的原发T1/T2 OSCC患者(n=32)和正常受试者(n=32)的非刺激性唾液。从唾液上清液中提取RNA，然后用T7RNA聚合酶进行两轮线性扩增。应用人类基因组U133A芯片分析人唾液转录组。采用t检验比较和折叠变化分析相结合的方法，对10例匹配的癌症患者和对照进行基因表达谱分析。采用定量聚合酶链式反应(QPCR)对筛选出的差异显著的基因进行芯片验证(P<；0.01)。结果：微阵列分析显示，有1,679个基因在癌症患者和对照组唾液中的表达水平存在显著差异(P<；0.05)。对口腔鳞癌患者(n=32)和对照组(n=32)唾液样本进行定量聚合酶链式反应(QPCR)，结果一致地证实了口腔鳞癌患者(n=32)和对照组(n=32)唾液中有7个与癌症相关的mRNA生物标记物的水平至少升高了3.5倍(P<；0.01)。这些潜在的唾液RNA生物标志物是IL8、IL1B、DUSP1、HA3、OAZ1、S100P和SAT的转录本。这些生物标志物的组合在区分口腔鳞癌和对照方面的敏感性(91%)和特异性(91%)。结论：唾液转录组诊断在口腔癌检测中的应用是成功的。这一新的临床方法可以被开发为一种强大的、高通量和可重复性的早期癌症检测工具。唾液转录组图谱可用于评估其在其他重大疾病应用以及正常健康监测中的有用性。
Purpose: Oral fluid (saliva) meets the demand for noninvasive, accessible, and highly efficient diagnostic medium. Recent discovery that a large panel of human RNA can be reliably detected in saliva gives rise to a novel clinical approach, salivary transcriptome diagnostics. The purpose of this study is to evaluate the diagnostic value of this new approach by using oral squamous cell carcinoma (OSCC) as the proof-of-principle disease.Experimental Design: Unstimulated saliva was collected from patients (n = 32) with primary T1/T2 OSCC and normal subjects (n = 32) with matched age, gender, and smoking history. RNA isolation was done from the saliva supernatant, followed by two-round linear amplification with T7 RNA polymerase. Human Genome U133A microarrays were applied for profiling human salivary transcriptome. The different gene expression patterns were analyzed by combining a t test comparison and a fold-change analysis on 10 matched cancer patients and controls. Quantitative polymerase chain reaction (qPCR) was used to validate the selected genes that showed significant difference (P < 0.01) by microarray. The predictive power of these salivary mRNA biomarkers was analyzed by receiver operating characteristic curve and classification models.Results: Microarray analysis showed there are 1,679 genes exhibited significantly different expression level in saliva between cancer patients and controls (P < 0.05). Seven cancer-related mRNA biomarkers that exhibited at least a 3.5-fold elevation in OSCC saliva (P < 0.01) were consistently validated by qPCR on saliva samples from OSCC patients (n = 32) and controls (n = 32). These potential salivary RNA biomarkers are transcripts of IL8, IL1B, DUSP1, HA3, OAZ1, S100P, and SAT. The combinations of these biomarkers yielded sensitivity (91%) and specificity (91%) in distinguishing OSCC from the controls.Conclusions: The utility of salivary transcriptome diagnostics is successfully demonstrated in this study for oral cancer detection. This novel clinical approach could be exploited to a robust, high-throughput, and reproducible tool for early cancer detection. Salivary transcriptome profiling can be applied to evaluate its usefulness for other major disease applications as well as for normal health surveillance.