Effects of hepatocyte growth factor on the expression of matrix metalloproteinases and their tissue inhibitors during the endometrial cancer invasion in a three-dimensional coculture

Abstract

Matrix metalloproteinase (MMP)-2 and -9 are secreted and translocated from endometrial stromal cells to HEC-1 A cells in a steroid-dependent manner. We investigated the paracrine effect of hepatocyte growth factor (HGF) on MMPs and metalloproteinase tissue inhibitor (TIMP) expression in stromal and endometrial cancer cells, and correlated with cancer cell invasiveness in three-dimensional (3D) coculture. The 3D coculture of endometrial stromal and cancer cell lines (HEC-1 A, HEC-IB, or KLE) were maintained in the presence or absence of HGF. The expression of MMP-2 and -9, MT1-MMP, TIMP-1 and -2 were examined by RT-PCR and zymography. Under the same conditions, invasion of the cancer cells was quantified by Boyden's chamber assay. HGF strongly induced MMP-9 mRNA expression in stromal cells, but had little effect on MMP-2 mRNA. MT1-MMP mRNA was detected only in KLE and stromal cells, which was also increased by HGF. TIMP-1 and -2 mRNAs was ubiquitous with no dependence on HGF. Zymographic analysis of MMPs showed that activation of MMP-2 and -9 was enhanced by HGF. A significant increase in invasion of all three cancer cells with HGF was observed. The effect of HGF on the invasiveness of 3D cocultured endometrial cancer cells and stromal cells appears to be due to induction of MMP-9 mRNA expression in stromal cells and /or increased activation of MMP-2 and MMP-9 by proteolytic digestion.

Matrix metalloproteinases in the pathophysiology and progression of gynecological malignancies: could their inhibition be an effective therapeutic approach?

“…The lack of MMP-2 or MMP-9 mRNA expression in cancer cells is consistent with other reports [64,68,69], suggesting that endometrial cancer cells utilize MMPs secreted by stromal cells in the invasion process.…”

“…Their data clearly demonstrate that the amount of active MMP-2 and MMP-9 increased with a corresponding decrease in the amount of pro-forms of MMP-2 and MMP-9 when HGF was present. This may be due to an induction in mRNA synthesis or promotion of conversion of pro-forms to active forms [68].…”

Modeling the early endometriotic lesion: mesothelium-endometrial cell co-culture increases endometrial invasion and alters mesothelial and endometrial gene transcription

“…c-Met, a proto-oncogene, is a tyrosine kinase receptor that is activated by its ligand, hepatocyte growth factor (HGF). Hepatocyte growth factor has been implicated in the pathogenesis of melanoma and breast, prostate, colon, pancreatic, hematologic, ovarian, and endometrial cancers (47)(48)(49). Other investigators have provided evidence suggesting that HGF interaction with c-Met plays a role in the pathogenesis of endometriosis.…”

Obesity, adipocytokines, and insulin resistance in breast cancer

“…A role for HGF in tumour cell invasion and metastasis was implied by its capacity for stimulating the breakdown of adherent junctions in epithelial cell colonies in vitro and the promotion of cell dispersal (113); hence its alternative descriptive name: 'scatter factor'. Subsequently, HGF was shown to stimulate tumour cell invasion by enhancing cell motility and the induction of protease expression (114)(115)(116). In a nude mouse model, human breast cancer cells transfected to express HGF showed an increased capacity for growth and metastasis by a process which involved both enhanced migration and tumour angiogenesis (117).…”