1997 Fiscal Year Final Research Report Summary
Production and characterization of newly developed anti-metastatic agents
| Project/Area Number |
07671774
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | HAMAMATSU UNIV.SCH.OF MED. |
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Principal Investigator |
KOBAYASHI Hiroshi Hamamatsu Univ.Sch.Med.Dept.of Obstet.Gynecol.Assist.Professor, 医学部附属病院, 講師 (40178330)
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| Co-Investigator(Kenkyū-buntansha) |
NISHIGUCHI Tomizo Hamamatsu Univ.Sch.Med.Dept.of Obstet.Gynecol.Assist.Professor, 医学部, 助手 (30198452)
TERAO Toshihiko Hamamatsu Univ.Sch.Med.Dept.of Obstet.Gynecol.Professor, 医学部, 教授 (60022852)
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| Project Period (FY) |
1995 – 1997
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| Keywords | ovarian cancer / invasion / metastasis |
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| Research Abstract |
Urinary trypsin inhibitor (UTI) efficiently inhibits tumor cell invasion and the formation of metastasis. The anti-metastatic effect is dependent on the COOH-terminal domain II of UTI (HI-8). In order to develop a molecule that binds with high affinity to the urokinase receptor (uPAR) on tumor cell surface, a bifunctional hybrid molecule (ATFHI) consisting of the uPAR-binding amino-terminal fragment (ATF) of uPA (amino acid sequence 1-134) at the NH_2-terminus of HI-8 was produced in Escherichia coli by genetic engineering. The purified hybrid protein inhibited trypsin and plasmin (inhibitory function was reduced two-to three-fold when compared with HI-8) and was also found to bind to human tumor cell via uPAR,which was confirmed by cell binding and competition experiments. Using a modified Boyden chamber and an artificial basement membrane, Matrigel, it was found that the hybrid protein is very effective at inhibiting invasion by uPAR-expressing human tumor cells. Sensitivities of tumor cells towards the anti-invasive effect of ATFHI correlated with the density of uPAR on human tumor cells. Furthermore, in the spontaneous metastasis model, the hybrid protein inhibited the formation of lung and /or lymphatic metastasis by human ovarian carcinoma and choriocarcinnoma cells. The hybrid protein was much more effective than ATF,HI-8, or UTI.We conclude that this approach extends the possibility of applying recombinant protein for therapeutic use in inhibition of human tumor cell metastasis.
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