Hepatocyte growth factor (HGF), a factor which stimulates cell growth, morphogenesis and migration, was found in hepatocytes and other epithelial cells. HGF was found to induce cell scattering, so that it was previously named as ¡§scatter factor¡¨ (SF), which turned out to be the same protein. HGF has two chains, a-chain, which contains N-terminal domain and four kringle domains and b-chain, a serine protease-like domain linked with a-chain by disulfide bond. The size of b-chain is 34 kDa after glycosylation in human cells. The function of HGFb was not unknown until recently that HGFb was shown 1000-fold lower affinity to c-Met than HGFa. Thus this project is to create biotechnological approaches for quick and large scale production of HGFb. The DNA fragment amplified by PCR was transferred to pET-22b(+) for expression, and showed that large amount of recombinant HGFb was produced both in periplasmic space and culture supernatant. Assays for cancer proliferation and migration with the HGFb showed that the recombinant protein inhibited cancer cells in a dose-independent manner. Our experiment showed clearly that 10 nM of the E. coli-produced HGFb inhibited proliferation and migration of cancer cells in comparison with the medium and pET controls, 1000-fold higher affinity to c-Met than reported.
| Identifer | oai:union.ndltd.org:NSYSU/oai:NSYSU:etd-0817107-133703 |
| Date | 17 August 2007 |
| Creators | Chen, Yi-Ting |
| Contributors | Shiping He, Ming-Hong Tai, Hung-Tu Huang |
| Publisher | NSYSU |
| Source Sets | NSYSU Electronic Thesis and Dissertation Archive |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0817107-133703 |
| Rights | withheld, Copyright information available at source archive |
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