Functhional analysis of tumor suppressur CHFR and development of novel daiagnotic and therapeutic strategies

• Project/Area Number: 22791293
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Digestive surgery
• Research Institution: Hokkaido University (2011); Sapporo Medical University (2010)
• Principal Investigator: KASHIMA Lisa 北海道大学, 大学院・薬学研究院, 博士研究員 (30404750)
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2011: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2010: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 癌 / CHFR / PARP-1 / 細胞周期チェックポイント / ユビキチン化 / タキサン系抗癌剤 / PARP阻害剤 / タキサン系抗がん剤

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 target 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) 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 prophase, supporting that the cells expressing CHFR were resistant to microtubule inhibitors. By contrast, in CHFR-silenced cells, polyubiquitination was not induced in response to mitotic stress. Thus, PARP-1 protein levels did not decrease, and cells progressed into mitosis under mitotic stress, suggesting that CHFR-silenced cancer cells were sensitized to microtubule inhibitors. Furthermore, we found that cells from Chfr knockout mice and CHFRsilenced 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.

Research Products

• [Journal Article] CHFR regulates the mitotic checkpoint by targeting PARP-1 for ubiquitination and degradation (2012)
  • Author(s): Kashima, L., Idogawa, M., Mita, H., Shitashige, M., Yamada, T., Ogi, K., Suzuki, H., Toyota, M., Ariga, H., Sasaki, Y., Tokino, T.
  • Journal Title: J. Biol. Chem Volume: 287 Issue: 16 Pages: 12975-12984
  • DOI: 10.1074/jbc.m111.321828
  • Peer Reviewed
• [Journal Article] Cystein-rich intestinal protein 2 (CRIP2) acts as a repressor of NFkB-mediated pro-angiogenic cytokins transcription to suppress tumorigenesis and angiogenesis (2011)
  • Author(s): Cheung, K.L., 7名略, Suzuki, T., 4名略
  • Journal Title: Proceeding of National Academy of Science USA Volume: 108 Issue: 20 Pages: 8390-8395
  • DOI: 10.1073/pnas.1101747108
  • Peer Reviewed
• [Journal Article] Identification and characterization of early growth response 2, a zinc-finger transcription factor, as a p53-regulated proapoptotic gene (2010)
  • Author(s): Yokota I, Sasaki Y, Kashima L, Idogawa M, Tokino T
  • Journal Title: International Journal of Oncology Volume: 37(6) Issue: 6 Pages: 1407-16
  • DOI: 10.3892/ijo_00000792
  • Peer Reviewed
• [Presentation] The functional relationship between CHFR and PARP-1 controls the antephase checkpoint and tumor development (2011)
  • Author(s): Kashima L
  • Organizer: 102nd AACR Annual Meeting
  • Place of Presentation: Orange County Convention Center (USA)
  • Year and Date: 2011-04-05
• [Presentation] CHFR regulates the mitotic checkpoint by targeting PARP-1 for ubiquitination and degradation (2011)
  • Author(s): Kashima L
  • Organizer: JSPS Sapporo Cancer Epigenetics Seminar of the A3 Foresight Program 2011, Session 3
  • Place of Presentation: New Otani in Sapporo(Hokkaido)
• [Presentation] The functional relationship between CHFR and PARP-1 controls the antephase checkpoint and tumor development (2011)
  • Author(s): Kashima L
  • Organizer: 102nd AACR Annual Meeting
  • Place of Presentation: Orenge County Convention Center(USA)
• [Presentation] Functional association between CHFR and PARP-1 controls the mitotic checkpoint. (2010)
  • Author(s): 鹿島理沙
  • Organizer: 日本癌学会総会
  • Place of Presentation: 大阪
  • Year and Date: 2010-09-23
• [Presentation] The functional relationship between CHFR and PARP-1 controls the mitotic checkpoint and tumor development (2010)
  • Author(s): 鹿島理沙
  • Organizer: 第33回日本分子生物学会年会
  • Place of Presentation: 神戸ポートアイランド(兵庫)
• [Presentation] Functional association between CHFR and PARP-1 controls the mitotic checkpoint (2010)
  • Author(s): 鹿島理沙
  • Organizer: 第69回日本癌学会総会
  • Place of Presentation: 大阪国際会議場(大阪)
• [Presentation] CHFR, a potent tumor suppressor, downregulates interleukin-8 via inhibition of NF-kappaB (2010)
  • Author(s): Lisa Kashima
  • Organizer: 8th Joint Conference of the American Association for Cancer Research and the Japanese Cancer Association
  • Place of Presentation: Hilton Waikoloa Village(USA)
• [Presentation] The Functional relationship between CHFR and PARP-1 controls the mitotic checkpoint and tumor development. (2010)
  • Author(s): 鹿島理沙
  • Organizer: 日本分子生物学会年会
  • Place of Presentation: 神戸(ポスター 口演)
• [Remarks] 研究の成果をホームページ上で随時、公開している
  • URL: http://web.sapmed.ac.jp/canmol/aisatsu.html