Areca quid chewing enhances the expression of salivary matrix metalloproteinase-9

Background and Purpose: The effects of areca quid (AQ) consumption on salivary matrix metalloproteinases (MMPs) which may participate in tumor invasion and metastasis remains unclear. This study assessed the change in salivary MMP-9 protein levels 2 hours after 5-minute AQ chewing stimulation (AQCS) in non-AQ users and the expression profile of this proteinase in saliva and tumor specimens of oral squamous cell carcinoma (OSCC) patients with a history of AQ use. Methods: MMP-9 transcript was analyzed by reverse transcription-polymerase chain reaction. MMP-9 protein level was measured by both Western blot and gelatin zymography. Results: The protein level of salivary MMP-9 was 3.1- to 8.9-fold enhanced 2 h after AQCS in 3 healthy volunteers as revealed by Western blot and zymography. As a control, gum chewing did not significantly change salivary MMP-9 protein level. Expression of MMP-9 transcript was found in 25 of 28 OSCC specimens and significantly correlated with cervical lymph node metastasis (p = 0.037). All of the 8 tested OSCC tissue homogenate samples available and all 12 saliva samples from 12 oral tumor outpatients were positive for MMP-9 protein. Conclusions: Elevation of MMP-9 may be one of the net effects of AQCS in vivo, which may play a role in the pathogenesis of oral mucosal lesions. Furthermore, the association of MMP-9 expression with neck-lymph-node metastasis may imply a significant role of MMP-9 in the progression of OSCC among patients with a history of AQ use in Taiwan.