title: Serum circulating human mRNA profiling and its utility for oral cancer detection.
abstract: OBJECTIVE
  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.
  
  
  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 > or = 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: School of Dentistry and Dental Research Institute, Division of Head & Neck Surgery/Otolaryngology, David Geffen School of Medicine University of California, Los Angeles, CA 90095, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.forename: David
authors.initials: D
authors.lastname: Elashoff
authors.forename: Myungshin
authors.initials: M
authors.lastname: Oh
authors.forename: Uttam
authors.initials: U
authors.lastname: Sinha
authors.forename: Maie A R
authors.initials: MA
authors.lastname: St John
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Zhou
authors.forename: Elliot
authors.initials: E
authors.lastname: Abemayor
authors.forename: David T
authors.initials: DT
authors.lastname: Wong
mesh_terms: D015415:Biomarkers / Q000097:blood; D002294:Carcinoma, Squamous Cell / Q000097:blood / Q000175:diagnosis / Q000235:genetics*; D016022:Case-Control Studies; D005260:Female; D006801:Humans; D016015:Logistic Models; D008297:Male; D046228:Microarray Analysis / Q000379:methods; D008875:Middle Aged; D009062:Mouth Neoplasms / Q000097:blood / Q000175:diagnosis / Q000235:genetics*; D012333:RNA, Messenger / Q000097:blood*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D015415:Biomarkers; D012333:RNA, Messenger