| Project/Area Number |
15592107
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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 |
Surgical dentistry
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| Research Institution | Osaka University |
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Principal Investigator |
SUMI Tetsuro Osaka University, Dental Hospital, Assistant Professor, 歯学部附属病院, 講師 (40252697)
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| Co-Investigator(Kenkyū-buntansha) |
YURA Yoshiaki Osaka University, Graduate School of Dentistry, Professor, 大学院・歯学研究科, 教授 (00136277)
NAKAZAWA Mitsuhiro Osaka University, Dental Hospital, Assistant Professor, 歯学部附属病院, 講師 (70217701)
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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 |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2003: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Teratocarcinoma / Cisplatin / p34^<cdc2> / リンパ管新生 / リンパ節転移 |
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| Research Abstract |
(1)F9 cells arrested at G21M with CDDP
Since the treatment with CDDP causes complete inhibition of growth in F9 cells, the effect of CDDP on cell cycle progression was analyzed using fiow cytometry. At various times after treatment with 5 x 10〜6 M CDDP, aliquots of cells were removed for flow cytometric analysis to determine the cell cycle stage. From these findings, the cell cycle was arrested at the late S + G2/M phase.
(2)Decrease in p34cdc2 by treatment with CDDP
The level of p34^<cdc2> in CDDP treated F9 cells was evaluated by Western blot analysis using anti- p34^<cdc2> antibody. Total cell lysates were prepared from cells collected at various times after treatment with CDDP. These lysates were assayed at equal cell numbers or at equal volumes of total protein. After the treatment with CDDP for 36 h, the level of p34^<cdc2> was decreased in the examination assay using equal cell numbers. In the assay using equal total proteins, a similar finding was observed after 36 h, but the level of p34^<cdc2> was increased for 24 h treatment with CDDP compared with the tTeatment for 0 h.
(3)The cdc2 mRNA was stable after the CDDP treatment
To clarify if the regulation of p34^<cdc2> by treatment with CDDP occurred in post- or pre-transcription, the level of cdc2 mRNA was estimated by Northern blotting analysis. By densitometric analysis, the level of cdc2 mRNA was stable for 36 h after treatment with CDDP.
(4)Turnover of p34^<cdc2>
To examine the half-life of p34^<cdc2> after treatment with CDDP, metabolic labeling and immunoprecipitation were carried out. The half-life of p34^<cdc2> in F9 cells treated with CDDP for 12 and 24 h was increased, but was decreased by treatment with CDDP for 36 h compared with control samples.
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