| Project/Area Number |
20390519
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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 |
Surgical dentistry
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| Research Institution | Sapporo Medical University |
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Principal Investigator |
TOKINO Takashi Sapporo Medical University, 医学部, 教授 (40202197)
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| Co-Investigator(Kenkyū-buntansha) |
HIRATSUKA Hiroyoshi 札幌医科大学, 医学部, 教授 (50165180)
OGI Kazuhiro 札幌医科大学, 医学部, 助教 (40433114)
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| Project Period (FY) |
2008 – 2010
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| Project Status |
Completed (Fiscal Year 2010)
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| Budget Amount *help |
¥19,240,000 (Direct Cost: ¥14,800,000、Indirect Cost: ¥4,440,000)
Fiscal Year 2010: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2009: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
Fiscal Year 2008: ¥7,670,000 (Direct Cost: ¥5,900,000、Indirect Cost: ¥1,770,000)
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| Keywords | 癌 / 歯学 / トランスレーショナルリサーチ / 細胞周期 / 微小管阻害剤 / CHFR / PARP-1阻害剤 / PARP-1 / トランスレーションリサーチ / 遺伝子 / ゲノム / 薬剤感受性 / ユビキチンリガーゼ / チェックポイント |
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| Research Abstract |
The mitotic checkpoint gene CHFR is silenced by promoter hypermethylation or mutated in various human cancers, suggesting that CHFR is an important tumor suppressor. Recent studies have reported that CHFR functions as an E3 ubiquitin ligase, resulting in the degradation of these proteins. To better understand how CHFR suppresses cell cycle progression and tumorigenesis, we sought to identify CHFR-interacting proteins using affinity purification combined with mass spectrometry. Herein, we showed poly(ADP-ribose) polymerase-1 (PARP-1), which catalyzes polyADP-ribosylation, to be a novel CHFR interacting protein. In CHFR expressing cells, mitotic stress induced the autoPARylation of PARP-1, resulting in an enhanced interaction between CHFR and PARP-1 and an increase in the polyubiquitination/degradation of PARP-1. The decrease in PARP-1 protein levels promoted cell cycle arrest at antephase, suggesting that the cells expressing CHFR were resistant to microtubule inhibitors. By contrast, in CHFR-silenced cells, polyubiquitination following autoPARylation of PARP-1 was not induced in response to mitotic stress. Thus, PARP-1 protein levels did not decrease, and cells progressed into mitosis under mitotic stress. Furthermore, we found that cells from Chfr knockout mice and CHFR-silenced primary gastric cancer tissues expressed higher levels of PARP-1 protein, strongly supporting our data that the interaction between CHFR and PARP-1 plays an important role in cell cycle regulation and cancer therapeutic strategies. Based on our studies, we demonstrate a significant advantage for use of combinational chemotherapy with PARP inhibitors for cancer cells resistant to microtubule inhibitors.
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