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Originally published as JCO Early Release 10.1200/JCO.2005.03.7598 on February 27 2006

Journal of Clinical Oncology, Vol 24, No 11 (April 10), 2006: pp. 1754-1760
© 2006 American Society of Clinical Oncology.

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Serum Circulating Human mRNA Profiling and Its Utility for Oral Cancer Detection

Yang Li, David Elashoff, Myungshin Oh, Uttam Sinha, Maie A.R. St John, Xiaofeng Zhou, Elliot Abemayor, David T. Wong

From the School of Dentistry and Dental Research Institute, Division of Head & Neck Surgery/Otolaryngology, David Geffen School of Medicine; Department of Biostatistics, School of Public Health, Henry Samueli School of Engineering and Applied Science; Jonsson Comprehensive Cancer Center and Molecular Biology Institute, University of California, Los Angeles (UCLA); and the School of Medicine, University of Southern California, Los Angeles, CA.

Address reprint requests to David T. Wong, DMD, DMSc, University of California Los Angeles, School of Dentistry, Dental Research Institute, 73-017 CHS, 10833 Le Conte Ave, Los Angeles, CA 90095; e-mail: dtww{at}ucla.edu

PURPOSE: The purpose of this study is to explore the presence of informative RNA biomarkers from human serum transcriptome, and evaluate the serum transcriptome diagnostics for disease detection. Oral squamous cell carcinoma (OSCC) was selected as the proof-of-concept disease.

PATIENTS AND METHODS: Blood samples were collected from patients (n = 32) with primary T1/T2 OSCC and matched healthy patients (n = 35). Circulating RNA was isolated from serum and linearly amplified using T7 polymerase. Microarrays were applied for profiling transcriptome in serum from 10 cancer patients and controls. The differential gene expression was analyzed by combining the present calls, t tests, and fold-change statistics. Quantitative polymerase chain reaction (PCR) was used to validate the selected candidate RNA markers identified by microarray. Receiver operating characteristic curve and classification models were exploited to evaluate the diagnostic power of these markers for OSCC.

RESULTS: Human serum circulating mRNAs were presented by reverse transcriptase PCR. Microarray identified 2,623 ± 868 probes assigned present calls in OSCC (n = 10) versus 1,792 ± 165 in healthy patients (n = 10), indicating a higher complexity of serum transciptome in OSCC patients (P = .002, Wilcoxon test). Three hundred thirty-five serum RNAs exhibited significantly differential expression level between the two groups (P < .05, t test; fold ≥ 2). Five cancer-related gene transcripts were consistently validated by quantitative PCR on serum from OSCC patients (n = 32) and controls (n = 35). The best combination of biomarkers yielded a receiver operating characteristic curve value of 88%, sensitivity (91%), and specificity (71%) in distinguishing OSCC.

CONCLUSION: The utility of serum transcriptome diagnostics is successfully demonstrated for OSCC detection. This novel concept could be developed as an adjunctive tool for disease diagnosis.

Supported by US Public Health Service (PHS) Grant No. RO1 DE15970, a UCLA Jonsson Comprehensive Cancer Center Grant (D.T.W.), PHS Grant No. T32 DE07296-07, and a Cancer Research Foundation of American fellowship (X.Z.).

Terms in blue are defined in the glossary, found at the end of this article and online at www.jco.org.

Authors' disclosures of potential conflicts of interest and author contributions are found at the end of this article.


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