| Project/Area Number |
15590094
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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 |
Biological pharmacy
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| Research Institution | Shujitsu University (2004)
National Cancer Center Research Institute and Research Center for Innovative Oncology, National Cancer Center Hospital East (2003) |
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Principal Investigator |
WATANABE Masahiko Shujitsu University, School of Pharmacy, Professor, 薬学部, 教授 (00182949)
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| Co-Investigator(Kenkyū-buntansha) |
WAKABAYASHI Keiji National Cancer Center Research Institute, Deputy Director, 副所長 (60158582)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2004: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2003: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | cabbage butterfly / pierisin / ADP-ribosylation / in vivo antitumor activity / HeLa cells / enzyme domain / acute toxicity |
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| Research Abstract |
Pierisin-1, a 98-kDa protein found in cabbage butterfly, exhibits cytotoxicity to cancer cells. To examine whether this protein has anti-cancer activity in viva, acute toxicity to mice and suppression effects to transplanted tumors were analyzed. The LD_<50> value for pierisin-1 was around 5 μg/kg body weight on intraperitoneal injection into BALB/c mice. BALB/c nu/nu female 6-week-old nude mice inoculated 1 x 10^7 HeLa cells on the first day were subjected for intraperitoneal injection of pierisin-1 at a dose of 3 μg/kg on the next day. Thereafter, animals were killed at day 80. Tumors were formed in all ten mice treated without pierisin-1 and the mean tumor weight was 1.16 g per mouse. In mice treated with pierisin-1, the mean tumor weight was 0.56 g per mouse, and its value was significantly lower and no tumors were found in three of ten mice tested. Our results indicate that pierisin-1 has antitumor activity in viva. Next, we purified 47.6 mg pierisin-1 from 1,340 pupae of Pieris rapae. Enzymatic characterization demonstrated that the maximum turnover number of the N-terminal polypeptide and trypsin-treated pierisin-1 were 55 and 25 per second, respectively. Inhibition of the N-terminal enzyme domain by the associated C-terminal fragment was evident under certain conditions. Long-term reactions indicated that a single molecule of pierisin-1 has the capacity to generate more than 10^6 ADP-ribosylated DNA adducts, which could cause the death of a mammalian cell.
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