Objective: The purpose of this study was to determine the effect of fibronectin on cell growth and activation of matrix metalloproteinase (MMPs), which related to aggressive behavior in human squamous carcinoma cell line-2 (HSC-2). Methods: HSC-2 was plated on uncoated and fibronectin-coated plate for 48 hours. Then, cell proliferation was assay by MTT, the conditioned medium was collected and proceeding to gelatin zymography. The level of MMP-2, MMP-9, MT1-MMP and TIMP-2 mRNA expression were analyzed by reverse transcription-polymerase chain reaction. Result: Fibronectin significantly increased the HSC-2 cell proliferation to 1.2-1.4 fold compared to uncoated plate. We could not observe the activation of both secreted MMP-2 and MMP-9. Conversely, the level of MMP-9 mRNA expression was decreased in fibronectin-coated plate. Moreover, there is no significantly change in the MT1-MMP and TIMP-2 mRNA expression. Conclusion: The present study provide the evidence that extracellular matrix, especially fibronectin, plays a role in modulating the cell growth and invasion of human squamous carcinoma cells. The interaction between HSC-2 and fibronectin enhance cell growth, whereas it has no effect on the activation of MMP-2 and MMP-9 activity. Down-regulation of MMP-9 mRNA expression by fibronectin further suggests the integrin signaling pathway on MMP-9 expression and tumor metastasis.