Ets-1 is an Ets family transcription factor, can up-regulate the transcription of matrix metalloproteinase (MMP) genes and confers an invasive phenotype on human cancer cells. HSC3 is an oral squamous cell carcinoma-derived cell line, and it manifests high levels of Ets-1 and MMP-9 gene expression that are associated with invasive potential. In this study, we investigated the effect of Ets-1 on the invasive properties of oral cancer from a molecular biological perspective. We constructed an Ets-1 antisense (AS) expression vector, transfected HSC3 cells with the vector, and obtained HSC3AS cells that express Ets-1 AS RNA. The expression of Ets-1 and MMP-9 was analyzed with RT-PCR. The invasive ability of the HSC3AS cells was determined using a matrigel invasion assay and MMP-9 production was measured using gelatin zymography. The amount of Ets-1 mRNA was significantly reduced in HSC3AS cells compared with parental HSC3 cells and the control transfected with empty vector. Matrigel invasion assay revealed that the HSC3AS cells had lower invasive ability. Gelatin zymography demonstrated that HSC3AS MMP-9 productions were decreased compared with those of parental HSC3 cells and the control. These results imply that transfection of AS Ets-1 inhibits oral cancer invasion by down-regulating MMP-9 genes.

Ets-1 is an Ets family transcription factor, can up-regulate the transcription of matrix metalloproteinase (MMP) genes and confers an invasive phenotype on human cancer cells. HSC3 is an oral squamous cell carcinoma-derived cell line, and it manifests high levels of Ets-1 and MMP-9 gene expression that are associated with invasive potential. In this study, we investigated the effect of Ets-1 on the invasive properties of oral cancer from a molecular biological perspective. We constructed an Ets-1 antisense (AS) expression vector, transfected HSC3 cells with the vector, and obtained HSC3AS cells that express Ets-1 AS RNA. The expression of Ets-1 and MMP-9 was analyzed with RT-PCR. The invasive ability of the HSC3AS cells was determined using a matrigel invasion assay and MMP-9 production was measured using gelatin zymography. The amount of Ets-1 mRNA was significantly reduced in HSC3AS cells compared with parental HSC3 cells and the control transfected with empty vector. Matrigel invasion assay revealed that the HSC3AS cells had lower invasive ability. Gelatin zymography demonstrated that HSC3AS MMP-9 productions were decreased compared with those of parental HSC3 cells and the control. These results imply that transfection of AS Ets-1 inhibits oral cancer invasion by down-regulating MMP-9 genes.

Heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF), which belongs to the EGF family, has been shown to stimulate the growth of a variety of cells in an autocrine or paracrine manner. Although HB-EGF is widely expressed in tumors when compared to normal tissue, its contribution to tumor invasion is still not known. In the present study, we analyzed the effects of HB-EGF on the invasion activity of a cultured oral cancer cell line using short interfering RNA (siRNA).Oral squamous carcinoma cell lines, HSC3 and SAS, were transfected with siRNA targeting HB-EGF. Expression of HB-EGF was analyzed by real-time PCR. The invasiveness of the transfected cells was determined using a matrigel invasion assay, and MMP-9 production was measured by RT-PCR and gelatin zymography.The expression of HB-EGF was reduced in HSC3-siRNA and SAS-siRNA cells. The matrigel invasion assay demonstrated that the invasiveness of HSC3-siRNA and SAS-siRNA cells was reduced. Gelatin zymography demonstrated that in HSC3-siRNA and SAS-siRNA cells, MMP9 production was decreased.These findings suggest that HB-EGF expression is related to the invasion activity of oral cancer, particularly via regulation of MMP9.