1988 Fiscal Year Final Research Report Summary
The basic research for antitumor effect of tumor necrosis facctor (TNF) against malignant brain-tumor
| Project/Area Number |
62570674
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Orthopaedic surgery
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| Research Institution | The University of Tsukuba |
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Principal Investigator |
NAKAGAWA Kunio Institute of Clinical Medicine, The University of Tsukuba, Assistant Professor, 臨床医学系, 助教授 (70114626)
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| Co-Investigator(Kenkyū-buntansha) |
吉川 靖三 筑波大学, 臨床医学系, 教授 (00010067)
大野 敦也 筑波大学, 臨床医学系, 講師 (50107645)
NOSE Tadao Institute of Clinical Medicine, The University of Tsukuba, Professor (10009699)
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| Project Period (FY) |
1987 – 1988
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| Keywords | Tumor necrosis factor / Malignant brain-tumor / 関節鏡 / 同種移植 |
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| Research Abstract |
The effect partially purified rat tumor necrosis factor (TNF) was tested against 9L rat brain tumor both in vivo and in vitro. The TNF-containing serum (TNS) was produced by intravenous injection of OK432 and lipopolysaccharide (LPS). Injection of TNS significantly (p<0.05) prolonged the survival time of brain tumor-bearing rats (29.9 12.6 days after tumor cell inoculation, as compared to 20.8 4.4 days in the untreated group).
In the in vitro assay, medium containing 50% TNS significantly decreased the viability of 9L brain tumor cells, by 57.6%, 50.0%, and 57.0% at 3, 5, and 7 days after the beginning of culture, respectively. TNS also displayed significant inhibition of cell growth, indicating a cytostatic effect. To verify TNS activity, TNS was partially purified by means of the DEAE-Sephadex a 50 batch ion exchange method and Sephadex G 200 column chromatography. Four fractions were tested in TNF-sensitive L(S) cells, TNF-resistant L(R) cells, and 9L brain tumor cells. Fraction 4 of TNS demonstrated 37.5%, 88.1%, and 43.2% cell viability against L(S), L(R), and 9L cells, respectively. On the other hand, fraction 4 of normal rat serum showed 87.5%, 87.8%, and 82.2% cell viability, respectively. These results strongly suggest the presence of TNF in the TNS produced by OK432 and LPS.
The effect of recombinant human TNF( -hTNF, kindly supplyed by Dainippon Pharmaceutical Co, LTD.) was also tseted against 9l brain tumor in vivo. When the 9L tumor reached to the 10mm in diameter, 5000unit of -hTNF with or without lymphokine activated killer cell was injected intraven ously or intratumorally. Fine injections were given to each mouse one week interval. The result was that the -htnf with LAK cells siginificantly prolonged their survival time (P<0.005) compared to other groups.
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