1996 Fiscal Year Final Research Report Summary
Novel Anti-Cancer Drug Composed of Human Proteins
| Project/Area Number |
07457262
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General surgery
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| Research Institution | Keio University |
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Principal Investigator |
UEDA Masakazu Keio Univ., School of Medicine Surgery, Assistante Professor, 医学部, 講師 (50142419)
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| Co-Investigator(Kenkyū-buntansha) |
TAKESHIMA Kaoru Keio Univ., School of Medicine Surgery, Assistant, 医学部, 助手 (50236460)
JINNO Hiromitsu Keio Univ., School of Medicine Surgery, Assistant, 医学部, 助手 (20216261)
KIKUCHI Kiyoshi Keio Univ., Cancer Center, Assistant, がんセンター, 助手 (30129407)
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| Project Period (FY) |
1995 – 1996
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| Keywords | Anti-cancer druo / EGF / RNase / immunotoxin / EGFreceptor |
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| Research Abstract |
1. Mammalian pancreatic robonuclease (RNase) was conjugated chemically via a disulfide bond to human epidermal growth factor (EGF). The human EGF-mam malian RNase conjugate had no cytotoxic effect against the EGFreceptor-deficient small-cell lung cancer cell line H69. But this conjugate showed dose-dependent cytotoxicity against squamous carcinoma cell lines and breast cancer cell lines and the cytotoxicity of the conjugate correlated positively with the level of expression of EGFR by each cell line. An unconjugated mixture of EGF and RNase had no greater effect than RNase alone on any cell lines.
2. Recombinant human RNase 1 (RNase1) was chemically linked to recombinant human EGF.The EGF-RNase conjugate showed dose-dependent cytotoxicity for EGF receptor-overexpressing cell lines. An unconjugated mixture of EGF and RNase had no greater effect than RNase alone. the conjugate showed no detectable cytotoxicity atainst EGF receptor-deficient small cell lung cancer cell.
3. The gene coding for human pancreatic RNase1, was fused with a gene coding for human EGF or human IL-2. These DNA were expressed in E.Coli, efficient amounts of the recombinant human protein were produced in inclusion bodies, solubilized, refolded in vitro, regained activity and purified to homogeneity.
4. The RNase-EGF conjugate was cytotoxic, EGF-receptor number dependently, to several squamous carcinoma cell lines. A mixture of free EGF and free RNase1 had no greater effect than RNase1 alone and no cytotoxicity was detectable for receptor deficient control cells.
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