Genomewide Study of Salivary MicroRNAs for Detection of Oral Cancer.
MicroRNAs (miRNAs) in human saliva have recently demonstrated to be potential biomarkers for diagnosis purposes. However, lack of well-characterized/matched clinical groups and lack of suitable endogenous control (EC) for salivary extracellular miRNA detection and normalization are among the restrictions of applying salivary-based miRNA biomarker discovery. In the present study, we examined the differential expression pattern of miRNAs among 4 groups of subjects-including patients with oral squamous cell carcinoma (OSCC), patients with OSCC in remission (OSCC-R), patients with oral lichen planus, and healthy controls (HCs)-using a genomewide high-throughput miRNA microarray. First, we systematically screened 10 pooling samples and 34 individual samples of different groups to find a proper EC miRNA. We then investigated the genomewide expression patterns of differentially expressed miRNAs in saliva of different groups using NanoString nCounter miRNA expression assay and real-time quantitative polymerase chain reaction, followed by construction of receiver operating characteristic curves to determine the sensitivity and specificity of the assay. We identified miRNA-191 as a suitable EC miRNA with minimal intergroup and intragroup variability, and we used it for normalization. Of more than 700 miRNAs tested, 13 were identified as being significantly deregulated in saliva of OSCC patients compared to HCs: 11 miRNAs were underexpressed (miRNA-136, miRNA-147, miRNA-1250, miRNA-148a, miRNA-632, miRNA-646, miRNA668, miRNA-877, miRNA-503, miRNA-220a, miRNA-323-5p), and 2 miRNAs were overexpressed (miRNA-24, miRNA-27b). MiRNA-136 was underexpressed in both OSCC vs. HCs and OSCC vs. OSCC-R. MiRNA-27b levels were significantly higher in OSCC patients compared to those found in HCs, patients with OSCC-R, and patients with oral lichen planus and served as a characteristic biomarker of OSCC. Receiver operating characteristic curve analyses showed that miRNA-27b could be a valuable biomarker for distinguishing OSCC patients from the other groups. Our novel findings established a reliable EC miRNA for salivary-based diagnostic and indicate that the salivary miRNA profiles are discriminatory in OSCC patients.
Department of Medicine, University of Massachusetts Medical School, Worcester, MA, USA Harvard Catalyst Laboratory for Innovative Translational Technologies, Harvard Medical School Boston, MA, USA f.m.heravi@gmail.com.
F
F
Momen-Heravi
Harvard Catalyst Laboratory for Innovative Translational Technologies, Harvard Medical School Boston, MA, USA.
A J
AJ
Trachtenberg
Harvard Catalyst Laboratory for Innovative Translational Technologies, Harvard Medical School Boston, MA, USA Department of Developmental Biology, Harvard School of Dental Medicine, Boston, MA, USA.
W P
WP
Kuo
Department of Diagnostic Sciences, Texas A&M University-Baylor College of Dentistry, Dallas, TX, USA.
Y S
YS
Cheng
D014408:Biomarkers, Tumor / Q000032:analysis*; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis* / Q000235:genetics; D055106:Genome-Wide Association Study; D006801:Humans; D017676:Lichen Planus, Oral / Q000175:diagnosis / Q000235:genetics; D035683:MicroRNAs / Q000032:analysis*; D046228:Microarray Analysis; D009062:Mouth Neoplasms / Q000175:diagnosis* / Q000235:genetics; D012372:ROC Curve; D060888:Real-Time Polymerase Chain Reaction; D012463:Saliva / Q000737:chemistry*; D012680:Sensitivity and Specificity
D014408:Biomarkers, Tumor; C577037:MIRN136 microRNA, human; C572377:MIRN147 microRNA, human; C550570:MIRN148 microRNA, human; C546177:MIRN191 microRNA, human; C543845:MIRN24 microRNA, human; C531388:MIRN27 microRNA, human; C545473:MIRN503 microRNA, human; C576158:MIRN632 microRNA, human; C000592812:MIRN646 microRNA, human; C570298:MIRN877 microRNA, human; D035683:MicroRNAs
biomarker; human; microRNA-191; microRNA-27a; saliva; squamous cell carcinoma