| Summary: | 博士 === 國立中山大學 === 生物科學系研究所 === 101 === Hepatocyte growth factor, a potent cytokine of mesenchymal origin exhibiting slews of physiological responses in the wide variety of cells like proliferation, migration and invasion upon binding to its receptor c-Met, but the mechanisms were not clearly understood where upon improper secretions of hepatocyte growth factor and activation of the c-Met results to the development of cancers. The human HGF variants are the natural occurring splice variants composed of N-terminal domain with their respective Kringle domains designated as K1, K2, K3 and K4. The biological properties of all the variants (NK1-NK4) of HGF have been controversial, and in-vitro studies suggest that these variants might have the therapeutic role as an HGF/SF antagonist. In order to clarify the NK-mediated inhibitory mechanism, we isolated and purified the NK variants of HGF (NK1-NK4) as the soluble form, from E.coli with GST as their tags. The topical application of these recombinant GST tagged HGF variant proteins (fusion proteins) on Human lung cancer cells (A549) Proliferation, migration, invasion and expression of different MMPs were assayed at 4nM concentration and compared to the medium control and second control GST (4nM). Our result demonstrated that these recombinant fusion NKs (GST with NKs) have significantly inhibited mitogen, motogen, morphogenesis of A549 at 4nM with downhill regulation of MMP-2 and MMP-9 and over expression of MMP-8. Therefore, our results suggest that in-vitro over expression of MMP-8 with downward regulation of MMP-9 can be considered as vital signals against metastasis of lung cancer cells, underlying the possible development of new therapeutic drug.
|
|---|
