We combined an iTRAQ-based quantitative proteomic analysis and the miRNA determination to profile potentially novel biomarker from oral cancer. There are 757 and 674 unique proteins identified from proteomic analysis, and 13 proteins displayed consistent underexpression (<0.67 fold) in normal tissues in comparison with the corresponding tumor tissues. After preliminary screening, EGFR, OAT, TPT1, ITGA6, G3BP1 and CB39L were the six genes validated in the 37 oral cancer patients (T1, n=10; T2, n=10; T3, n=10 and T4, n=7). The TPT1, ITGA6 and CAB39L genes were displayed the higher transcriptions level in the tumor tissues and the TPT1, ITGA6 and CAB39L proteins were also shown overexpression in the tumor tissues from the same patients. The miR-19a, 19b, 27a, 27b, 186, 203 and 377 transcripts were predicted and the miR-27b level was shown to significantly reduce in the tumor tissues and the plasma of OSCC patients. In the in vitro study, the overexpression of miR-27b only significantly decreased TCTP protein and gene levels in both HSC-3 and Cal-27 cell lines. Our results demonstrate that human miR-27b regulates the expression of the TCTP tumor protein, and circulating miR-27b may be useful as a biomarker for oral cancer research.