Hepatocyte growth factor upregulates E1AF that induces oral squamous cell carcinoma cell invasion by activating matrix metalloproteinase genes

Abstract

Hepatocyte growth factor (HGF) is thought to play a role in cell motility and invasion. Matrix metalloproteinases (MMPs) have been implicated in invasion and metastasis of tumor cells. We have previously reported that the Ets-oncogene family transcription factor E1AF positively regulates transcription of MMP genes in transient expression assays and that overexpression of the E1AF gene confers an invasive phenotype on breast cancer cells. Here we examined the effect of HGF on E1AF and MMP gene expression in terms of the invasive potential of the oral squamous cell carcinoma cell line HSC3. HGF stimulated expression of the E1AF gene. The levels of MMP-1, -3 and -9 mRNAs increased in cells treated with HGF and correlated with E1AF upregulation. In contrast, no obvious upregulation of MMP-1 and -9 mRNA was observed in ASE1AFHSC3 cells transfected with the antisense E1AF expression vector into parental HSC3 cells. The wild-type MMP-9 gene promoter was activated by endogenous E1AF in HSC3 cells, and chloramphenicol acetyltransferase (CAT) activities increased when HGF was added to transfected cells. On the other hand, CAT activity was reduced to almost two-thirds of the wild-type activity when HSC3 cells were transfected with a CAT reporter plasmid driven by a mutant MMP-9 promoter lacking the Ets-binding site, and induction of CAT activity was not observed upon addition of HGF. Analysis of organotypic raft cultures revealed that HSC3 cells invaded and degraded collagen gel actively upon addition of HGF. These results suggest that HGF induces expression of the Ets-related E1AF transcription factor gene whose product in turn activates MMP genes and leads to oral cancer cell invasion.

Regulation of microRNA expression by hepatocyte growth factor in human head and neck squamous cell carcinoma

“…(5)(6)(7) Indeed, it has been reported that HGF ⁄ SF promotes cell migration and angiogenesis in human esophageal SCC, (27) upregulates the expression of proangiogenic cytokines interleukin (IL)-8 and vascular endothelial growth factor (VEGF) through the activation of mitogen-activated protein ⁄ extracellular signal-regulated kinase kinase (MEK) and phosphatidylinositol 3¢-kinase (PI3K) signaling pathways in HNSCC, (28) and induces the invasion of oral SCC by matrix metalloproteinase genes through the upregulation of the transcription factor E1AF. (29) However, how HGF ⁄ SF and MET signaling cascade affects the expression of each specific downstream functional gene has not yet been elucidated in detail. In addition, there is a growing number of reports that describe the translational regulation of growth factors or their receptors, including HGF and MET, in cancer by miR-NA, (30,31) but less reports describe the regulation of miRNA expression by a growth factor itself to date.…”

Role of HGF/c-met system in invasion and metastasis of oral squamous cell carcinoma cellsin vitro and its clinical significance

“…Moreover, several investigators reported that HGF up-regulated matrix-degrading enzymes. [31][32][33][34] We also reported that the augmentation of HGF/c-met signal upregulated proMMP9, proMMP1, and uPA in a renal cell carcinoma cell line, Caki-1. 18 In this oral SCC cell system, we also examined the effect of HGF on the activity of the gelatinase produced by BHY and HN cells.…”

“…Taq DNA polymerase (Takara Biomedicals, Kusatsu, Japan) was added to the mixture at a final concentration of 0.05 U/ l, and the reaction was carried out in a Takara Thermal Cycler MP (Takara Biomedicals) under the following conditions: 94°C for 3 min and then 94°C for 1 min, 55°C for 1.5 min, 72°C for 2.5 min for 30 cycles, and extension at 72°C for 4 min. To determine whether conditions were adequate for semi-quantitative RT-PCR, we reverse-transcribed 0.05, 0.5, 5, and 10 g of total RNA and amplified the fragments under the same PCR conditions with different numbers of cycles (18,22,26,30,34 cycles). We used 5 g of cytoplasmic or total RNA as a template for reverse transcription and 30 cycles of PCR amplification for c-met and 22 cycles for GAPDH.…”

Transcription factors control invasion: AP-1 the first among equals

“…Antisense oligonucleotides to Ets-1 reduced invasion in oral cancers. In the same system, HGF induced invasion was partially dependent upon PEA3/E1AF mediated induction of MMP9 (Hanzawa et al, 2000). Dominant-negative constructs consisting of the Ets domain of Ets-2 inhibit anchorage independent growth and invasion of BT20 breast carcinoma cells (Sapi et al, 1998) and PPC-1 prostate carcinoma cells (Foos and Hauser, 2000).…”