Purpose: We have previously shown that a transcriptome is found in saliva and subpanels of these mRNAs can be used as oral cancer biomarkers. In this study, we measured the presence of microRNAs (miRNA) in saliva and determined their potential as an additional set of oral cancer biomarkers. Experimental Design: A total of 314 miRNAs were measured using reverse transcriptase-preamplification-quantitative PCR in 12 healthy controls. Degradation pattern of endogenous and exogenous saliva miRNAs were measured at room temperature over time. Selected miRNAs were validated in saliva of 50 oral squamous cell carcinoma patients and 50 healthy matched control subjects. Results: We detected ∼50 miRNAs in both the whole and supernatant saliva. Endogenous saliva miRNA degraded much slower compared with exogenous miRNA. Two miRNAs, miR-125a and miR-200a, were present in significantly lower levels (P < 0.05) in the saliva of oral squamous cell carcinoma patients than in control subjects. Conclusions: Both whole and supernatant saliva of healthy controls contained dozens of miRNAs, and similar to saliva mRNAs, these miRNAs are stable. Saliva miRNAs can be used for oral cancer detection. (Clin Cancer Res 2009;15(17):5473-7)

Background Oral cancer is the sixth most common cancer with a five-year survival rate of approximately 60%. Presently there are no scientifically credible early detection techniques beyond conventional clinical oral examination. The goal of this study is to validate if the 7 mRNAs and 3 proteins previously reported biomarkers are capable of discriminating patients with oral squamous cell carcinomas (OSCC) from healthy subjects in independent cohorts and by a National Cancer Institute (NCI)- Early Detection Research Network (EDRN) Biomarker Reference Laboratory (BRL). Methods 395 subjects from 5 independent cohorts based on case-controlled design were investigated by 2 independent laboratories, UCLA discovery laboratory and NCI-EDRN Biomarker Reference Laboratory (BRL). Results Expression of all 7 mRNA and 3 protein markers was increased in OSCC versus controls in all 5 cohorts. With respect to individual marker performance across the 5 cohorts, the increase in IL-8 and SAT were statistically significant and remained top performers across different cohorts in terms of sensitivity and specificity. A previously identified multiple marker model demonstrated an area under the receiver operating characteristic (ROC)-curve for prediction of OSCC status ranging from of 0.74 to 0.86 across the cohorts. Conclusions The validation of these biomarkers demonstrated their feasibility in the discrimination of OSCC from healthy controls. Established assay technologies are robust enough to perform independently. Individual cutoff values for each of these markers and for the combined predictive model need to be further defined in large clinical studies. Impact Salivary proteomic and transcriptomic biomarkers can discriminate oral cancer from control subjects.

Early detection of oral squamous cell cancer (OSCC) is the key to improve the low 5-year survival rate. Using proteomic and genomic technologies we have previously discovered and validated salivary OSCC markers in American patients. The question arises whether these biomarkers are discriminatory in cohorts of different ethnic background. Six transcriptome (DUSP1, IL8, IL1B, OAZ1, SAT1, S100P) and three proteome (IL1B, IL8, M2BP) biomarkers were tested on 18 early and 17 late stage OSCC patients and 51 healthy controls with quantitative PCR and ELISA. Four transcriptome (IL8, IL1B, SAT1, S100P) and all proteome biomarkers were significantly elevated (p<0.05) in OSCC patients. The combination of markers yielded an AUC of 0.86, 0.85 and 0.88 for OSCC total, T1-T2, and T3-T4 respectively. The sensitivity/specificity for OSCC total was Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access Author ManuscriptOral Oncol. Author manuscript; available in PMC 2012 January 1. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript 0.89/0.78, for T1-T2 0.67/0.96, and for T3-T4 0.82/0.84. In conclusion, seven of the nine salivary biomarkers (3 proteins and 4 mRNAs) were validated and performed strongest in late stage cancer. Patient-based salivary diagnostics is a highly promising approach for OSCC detection. This study shows that previously discovered and validated salivary OSCC biomarkers are discriminatory and reproducible in a different ethnic cohort. These findings support the feasibility to implement multi-center, multi-ethnicity clinical trials towards the pivotal validation of salivary biomarkers for OSCC detection.

The Adult Comorbidity Evaluation 27 index is a more reliable predictor of survival than the Charlson Comorbidity Index in patients with laryngeal squamous cell carcinoma.