Identification of a new tumor suppressor protein involved in apoptosis and investigation of new treatment using the new tumor suppressor

• Project/Area Number: 18591824
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Obstetrics and gynecology
• Research Institution: The University of Tokyo
• Principal Investigator: NAKAGAWA Shunsuke The University of Tokyo, Faculty of Medicine, Lecturer (70270874)
• Co-Investigator(Kenkyū-buntansha): YANO Tetsu The University of Tokyo, Faculty of Medicine, Associate Professor (90251264)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥3,890,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥390,000); Fiscal Year 2007: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2006: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: Apoptosis / tumor suppressor / human papillomavirus / ubiquitination / E6 oncoprotein / 新規癌抑制遺伝子 / 子宮頸部腺癌

Research Abstract

Inactivation of tumor suppressor proteins involving in cell cycle regulation and apoptosis through ubiquitin-mediated degradation by human papillomavirus (HPV) E6 oncoprotein and ubiquitin-proteasome is a critical step in cervical carcinogenesis. Here, we show that HPV E6 targets the product of dbc-1 (deleted in breast cancer-1) gene, which was identified from homozygously deleted region in breast cancers on human chromosome 8p21, for ubiquitin-mediated degradation. DBC-1 interacts with both low- and high-risk HPV E6s, but its binding ability with low-risk E6 was weaker. Mutational analysis showed that E6 binds 700-750 amino acids region of DBC-1. In vitro translated DBC-1 with rabbit reticulocyte lysate but not that translated with wheat germ extract was degraded in the presence of high-risk E6. HPV 16 E6 stimulated in vitro ubiquitination of DBC-1 in the presence of ATP γ S. Transfection of high-risk E6 expression vector into 293 T cells induces ubiquitin-mediated degradation of DBC-1. Addition of proteasomal inhibitor MG132 or RNAi against E6AP recovered the DBC-1 expression in HPV-positive Caski cells. DBC-1 localized in nucleus in healthy cells, but concentrated in mitochondria during apoptosis induced by etoposide. Progressive loss of DBC-1 expression is observed during process of cervical carcinogenesis from normal cervical epithelia to invasive cervical cancer. These data suggest the possibility that degradation of DBC-1 by E6-E6AP might have a critical role in inhibition of apoptotic signal pathway in mitochondria during cervical carcinogenesis.

Research Products

• [Journal Article] hScrib, a human homologue of Drosophila neoplastic tumor suppressor, is a novel death substrate targeted by caspase during the process of apoptosis. (2008)
  • Author(s): Tsuruga T, Nakagawa S, et. al.
  • Journal Title: Genes to Cells. (In press)
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Cellular mechanisms of growth inhibition of human endometrial cancer cell line by an antagonist of growth hormone-releasing hormone. (2008)
  • Author(s): Zhao L, Yano T, Osuga Y, Nakagawa S., et. al.
  • Journal Title: Int J Oncol. 32(3) Pages: 593-601
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Cellular mechanisms of growth inhibition of human endometrial cancer cell line by an antagonist of growth hormone-releasing hormone (2008)
  • Author(s): Zhao L, Yano T, Osuga Y, Nakagawa S, Oishi H, Wada-Hiraike O, Tang X, Yano N, Kugu K, Schally AV, Taketani Y.
  • Journal Title: Int J Oncol 32(3) Pages: 593-601
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Combined phospho-Akt and PTEN expressions associated with post-treatment hysterectomy after conservative progestin therapy in complex atypical hyperplasia and stage la, G1 adenocarcinoma of the endometrium (2008)
  • Author(s): Minaguchi T, Nakagawa S, Takazawa Y, Nei T, Horie K, Fujiwara T, Osuga Y, Yasugi T, Kugu K, Yano T, Yoshikawa H, Taketani Y.
  • Journal Title: Cancer Lett 248(1) Pages: 112-22
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cellular mechanisms of growth inhibition of human endometrial cancer cell line by anantagonist of growth hormone-releasing hormone. (2008)
  • Author(s): Zhao L, Yano T, Osuga Y, Naka, gawa S, et. al.
  • Journal Title: Int J oncol 32(3) Pages: 593-60
  • Peer Reviewed
• [Journal Article] hScrib, a human homologue of Drosophila neoplastic tumor suppressor, is a novel death substrate targeted by caspase during process of apoptosis (2008)
  • Author(s): Sone K, Nakagawa S, et. al.
  • Journal Title: Genes to Cells 掲載確定
  • Peer Reviewed
• [Journal Article] Loss of Hugl-1 expression associates with lymph node metastasis in endometrial cancer. (2007)
  • Author(s): Tsuruga T, Nakagawa S, et. al.
  • Journal Title: Oncol Res. 16(9) Pages: 431-5
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Treatment with paclitaxel plus carboplatin, alone or with irradiation, of advanced or recurrent endometrial carcinoma. (2007)
  • Author(s): Arimoto T, Nakagawa S, et. al.
  • Journal Title: Gynecol Oncol. 104(1) Pages: 32-5
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Combined phospho-Akt and PTEN expressions associated with post-treatment hysterectomy after conservative progestin therapy in complex atypical hyperplasia and stage Ia, G1 adenocarcinoma of the endometrium. (2007)
  • Author(s): Minaguchi T, Nakagawa S, et. al.
  • Journal Title: Cancer Lett 8;248(1) Pages: 112-22
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Loss of Hugl-1 expression associates with lymph node metastasis in endometrial cancer (2007)
  • Author(s): Tsuruga T, Nakagawa S, Watanabe M, Takizawa S, Matsumoto Y, Nagasaka K, Sane K, Hiraike H, Miyamoto Y, Hiraike O, Minaguchi T, Oda K, Yasugi T, Yano T, Taketani Y.
  • Journal Title: Oncol Res 16(9) Pages: 431-5
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Treatment with paclitaxel plus carboplatin, alone or with irradiation, of advanced or recurrent endometrial carcinoma (2007)
  • Author(s): Arimoto T, Nakagawa S, Yasugi T, Yoshikawa H, Kawana K, Yano T, Taketani Y.
  • Journal Title: Gynecol Oncol 104(1) Pages: 32-5
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Treatment with pacliaxel plus carboplatin, alone or with irradiation, of advanced or recurrent endometrial carcinoma. (2007)
  • Author(s): Arimoto T, Nakagawa S, et. al.
  • Journal Title: Gynecol Oncol. 104(1) Pages: 32-35
  • Peer Reviewed
• [Journal Article] Combined phospho-Akt and PTEN expressions associated with post-treatment hysterectomy after conservative progestin therapy in complex atypical hyperplasia and stage Ia, G1 adenocarcinoma of the endometrium (2007)
  • Author(s): Minaguchi T, Nakagawa S, Takazawa Y, Nei T, Horie K, Fujiwara T, Osuga Y, Yasugi T, Kugu K, Yano T, Yoshikawa H, Taketani Y.
  • Journal Title: Cancer Lett. 248(1) Pages: 112-122
• [Journal Article] Human homolog of Drosophila tumor suppressor Scribble negatively regulates cell-cycle progression from G1 to S phase by localizing at the basolateral membrane in epithelial cells. (2006)
  • Author(s): Nagasaka K, Nakagawa S
  • Journal Title: Cancer Sci. 1217-25. 97(11) Pages: 1217-25
  • NAID: 10019377176
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Human scribble, a novel tumor suppressor identified as a target of high-risk HPV'E6 for ubiquitin-mediated degradation, interacts with adenomatous polyposis coli. (2006)
  • Author(s): Takizawa S, Nagasaka K, Nakagawa S, Et al.
  • Journal Title: Genes Cells. 11(4) Pages: 453-64
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Involvement of a cellular ubiquitin-protein ligase E6AP in the ubiquitin-mediated degradation of extensive substrates of high-risk human papillomavirus E6. (2006)
  • Author(s): Matsumoto Y, Nakagawa S, et. al.
  • Journal Title: J Med Virol. 78(4) Pages: 501-7
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Human homolog of Drosophila tumor suppressor Scribble negatively regulates cell-cycle progression from G1 to S phase by localizing at the basolateral membrane in epithelial cells (2006)
  • Author(s): Nagasaka K, Nakagawa S, Yano T, Takizawa S, Matsumoto Y, Tsuruga T, Nakagawa K, Minaguchi T, Oda K, Hiraike-Wada O, Ooishi H, Yasugi T, Taketani Y.
  • Journal Title: Cancer Sci 97(11) Pages: 1217-25
  • NAID: 10019377176
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Human scribble, a novel tumor suppressor identified as a target of high-risk HPV E6 for ubiquitin-mediated degradation, interacts with adenomatous polyposis coli (2006)
  • Author(s): Takizawa S, Nagasaka K, Nakagawa S, Yano T, Nakagawa K, Yasugi T, Takeuchi T, Kanda T, Huibregtse JM, Akiyama T, Taketani Y.
  • Journal Title: Genes Cells 11(4) Pages: 453-64
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Involvement of a cellular ubiquitin-protein ligase E6AP in the ubiquitin-mediated degradation of extensive substrates of high-risk human papillomavirus E6 (2006)
  • Author(s): Matsumoto Y, Nakagawa S, Yano T, Takizawa S, Nagasaka K, Nakagawa K, Minaguchi T, Wada O, Ooishi H, Matsumoto K, Yasugi T, Kanda T, Huibregtse JM, Taketani Y.
  • Journal Title: J Med Virol 78(4) Pages: 501-7
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Human Scribble, a novel tumor suppressor identified as a target of high-risk HPV E6 for ubiquitin-mediated degradation, interacts with adenomatous polyposis coli (2006)
  • Author(s): Takizawa S, Nagasaka K, Nakagawa S, Yano T, Matsumoto Y, Nakagawa K, Minaguchi T, Oda K, Hiraike-Wada O, Ooishi H, Matsumoto K, Yasugi T, Taketani Y
  • Journal Title: Genes to Cells. 2006 11 Pages: 453-464
• [Journal Article] Involvement of a cellular ubiquitin-protein ligase E6AP in the ubiquitin-mediated degradation of extensive substrates of high-risk human papillomavirus E6. (2006)
  • Author(s): Matsumoto Y, Nakagawa S, Yano T, Takizawa S, Nagasaka K, Nakagawa K, Minaguchi T, Wada O, Ooishi H, Matsuiuoto K, Yasugi T, Kanda T, Huibregtse JM, Taketani Y.
  • Journal Title: J Med Virol. 78(4) Pages: 501-507
• [Journal Article] Human scribble, a novel tumor suppressor identified as a target of high-risk HPV E6 for ubiquitin-mediated degradation, interacts with adenomatous polyposis coli (2006)
  • Author(s): Takizawa S, Nagasaka K, Nakagawa S, Yano T, Nakagawa K, Yasugi T, Takeuchi T, Kanda T, Huibregtse JM, Akiyama T, Taketani Y.
  • Journal Title: Genes to Cells 11(4) Pages: 453-464
• [Journal Article] Human homolog of Drosophila tumor suppressor Scribble negatively regulates cell-cycle progression from G1 to S phase by localizing at the basolateral membrane in epithelial cells (2006)
  • Author(s): Nagasaka K, Nakagawa S, Yano T, Takizawa S, Matsumoto Y, Tsuruga T, Nakagawa K, Minaguchi T, Oda K, Hiraike-Wada O, Ooishi H, Yasugi T, Taketani Y.
  • Journal Title: Cancer Sci. 97(11) Pages: 1217-1225
  • NAID: 10019377176
• [Journal Article] hScrib, a human homologue of Drosophila neoplastic tumor suppressor, is a novel death substrate targeted by caspase during the process of apoptosis
  • Author(s): Kenbun Sone, Shunsuke Nakagawa
  • Journal Title: Genes to Cells (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Analysis of the expression and localisation of a LAP protein, human scribble, in the normal and neoplastic epithelium of uterine cervix.
  • Author(s): Nakagawa S., Yano T, Nakagawa K, Takizawa S, Suzuki Y, Yasugi T, Huibregtse JM, Taketani Y.
  • Journal Title: Br J Cancer.
• [Journal Article] The DNA mismatch repair gene hMSH2 is a potent coactivator of oestrogen receptor alpha
  • Author(s): Wada-Hiraike O, Yano T, Nei T, Matsumoto Y, Nagasaka K, Takizawa S, Oishi H, Arimoto T, Nakagawa S, Yasugi T, Kato S, Taketani Y.
  • Journal Title: Br J Cancer. 2005 Pages: 2286-2291
• [Journal Article] Comparison between conventional surgery and radiotherapy for FIGO stage I-II cervical carcinoma : a retrospective Japanese study
  • Author(s): Yamashita H, Nakagawa K, Tago M, Shiraishi K, Nakamura N, Ohtomo K, Oda K, Nakagawa S., Yasugi T, Taketani Y.
  • Journal Title: Gynecol Oncol. Pages: 834-839
• [Presentation] hScrib, a human homologue of Drosophila neoplastic tumor suppressor, is a novel death substrate targeted by caspase during process of apoptosis (2007)
  • Author(s): Sone K, Nakagawa S, et. al.
  • Organizer: 日本癌学会
  • Place of Presentation: 横浜
  • Year and Date: 2007-10-27
• [Presentation] Deleted in breast cancer-1 (DBC-1) is targeted for the 2007 July, ubiquitie-mediated degradation by human papillomavirus E6 oncoprotein (2007)
  • Author(s): Yoko Matsumoto
  • Organizer: DNA tumor virus
  • Place of Presentation: Trieste, Italia
  • Year and Date: 2007-07-21
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Deleted in breast cancer-1(DBC-1) is targeted for the ubiquitin-mediated degradation by human papillomavirus E6 oncoprotein (2007)
  • Author(s): Yoko Matsumoto
  • Organizer: DNA tumor virus
  • Place of Presentation: Trieste, Italia
  • Description: 「研究成果報告書概要(欧文)」より