Functional analysis of CHFR: Diagnostic and therapeutic application for oral squamous cell cancer

2010 Fiscal Year Final Research Report

• Project/Area Number: 20390519
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Surgical dentistry
• Research Institution: Sapporo Medical University
• Principal Investigator: TOKINO Takashi Sapporo Medical University, 医学部, 教授 (40202197)
• 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)
• Project Period (FY): 2008 – 2010
• 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

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.

Research Products

• [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: Int J Oncology Vol.37, No.6 Pages: 1407-1416
  • Peer Reviewed
• [Journal Article] Genomic screening for genes upregulated by demethylation revealed novel targets of epigenetic silencing in breast cancer. (2010)
  • Author(s): Fujikane T, Nishikawa N, Toyota M, Suzuki H, Nojima M, Maruyama R, Ashida M, Ohe-Toyota M, Kai M, Nishidate T, Sasaki Y, Ohmura T, Hirata K, Tokino T
  • Journal Title: Breast Cancer Res Treat Vol.122, No.3 Pages: 699-710
  • Peer Reviewed
• [Journal Article] A novel correlation between LINE-1 hypomethylation and the malignancy of gastrointesrinal stromal tumors. (2010)
  • Author(s): Igarashi S, Suzuki H, Niinuma T, Nojima M, Iwaki H, Nobuoka T, Nishida T, Miyazaki Y, Takamaru H, Yamamoto E, Yamamoto H, Tokino T, Hasegawa T, Hirata K, Imai K, Toyota M, Shinomura Y
  • Journal Title: Clin Cancer Res Vol.16, No.21 Pages: 5114-5123
  • Peer Reviewed
• [Journal Article] IGFBP7 is a p53 responsive gene specifically silenced in colorectal cancer with CpG island methylator phenotype. (2010)
  • Author(s): Suzuki H, Igarashi S, Nojima M, Maruyama R, Yamamoto E, Kai M, Akashi H, Watanabe Y, Yamamoto H, Sasaki Y, Itoh F, Imai K, Sugai T, Shen L, Issa J-P J, Shinomura Y, Tokino T, Toyota M
  • Journal Title: Carcinogenesis Vol.31, No.3 Pages: 342-349
  • Peer Reviewed
• [Journal Article] DNA methylation of interferon regulatory factors in gastric cancer and noncancerous gastric mucosae. (2010)
  • Author(s): Yamashita M, Toyota M, Suzuki H, Nojima M, Yamamoto E, Kamimae S, Watanabe Y, Kai M, Akashi H, Maruyama R, Sasaki Y, Yamano H, Sugai T, Shinomura Y, Imai K, Tokino T, Itoh F
  • Journal Title: Cancer Sci Vol.101, No.7 Pages: 1708-1716
  • Peer Reviewed
• [Journal Article] CHFR, a potential tumor suppressor, downregulates interleukin-8 via inhibition of NF-kB. (2009)
  • Author(s): Kashima L, Toyota M, Mita H, Suzuki H, Idogawa M, Ogi K, Sasaki Y, Tokino T
  • Journal Title: Oncogene Vol.28, No.29 Pages: 2643-2653
  • Peer Reviewed
• [Journal Article] A single recombinant adenovirus expressing p53 and p21-targeting artificial microRNAs efficiently induces apoptosis in human cancer cells. (2009)
  • Author(s): Idogawa M, Sasaki Y, Suzuki H, Mita H, Imai K, Shinomura Y, Tokino T
  • Journal Title: Clin Cancer Res Vol.15, No.11 Pages: 3725-3732
  • Peer Reviewed
• [Journal Article] p53 family members regulate the expression of the apolipoprotein D gene. (2009)
  • Author(s): Sasaki Y, Negishi H, Koyama R, Anbo N, Ohori K, Idogawa M, Mita H, Toyota M, Imai K, Shinomura Y, Tokino T
  • Journal Title: J Biol Chem Vol.284, No.2 Pages: 872-883
  • Peer Reviewed
• [Journal Article] A novel method, digital genome scanning detects KRAS gene amplification in gastric cancers : involvement of overexpressed wild-type KRAS in downstream signaling and cancer cell growth. (2009)
  • Author(s): Mita H, Toyota M, Aoki F, Akashi H, Maruyama R, Sasaki Y, Suzuki H, Idogawa M, Kashima L, Yanagihara K, Fujita M, Hosokawa M, Kusano M, Sabau SV, Tatsumi H, Imai K, Shinomura Y, Tokino T
  • Journal Title: BMC Cancer Vol.9, No.198(on line)
  • Peer Reviewed
• [Journal Article] A novel biomarker for cervical cancer. (2009)
  • Author(s): Tokino T
  • Journal Title: Cancer Biol Ther Vol.8, No.12 Pages: 1154-1155
  • Peer Reviewed
• [Journal Article] Genomic Screening for genes silenced by DNA methylation revealed an association between RASD1 inactivation and dexamethasone resistence in multiple myeloma. (2009)
  • Author(s): Nojima M, Maruyama R, Yasui H, Suzuki H,Maruyama Y, Tarasawa I, Sasaki Y, Asaoku H, Sakai H, Hayashi T, Mori M, Imai K, Tokino T, Ishida T, Toyota M, Shinomura Y
  • Journal Title: Clin Cancer Res1 Vol.15, No.13 Pages: 4356-4364
  • Peer Reviewed
• [Journal Article] Overexpression of Aurora A by loss of CHFR gene expression increases the growth and survival of HTLV-1-infected T cells through enhanced NF-kappaB activity. (2009)
  • Author(s): Tomita M, Toyota M, Ishikawa C, Nakazato T, Okudaira T, Matsuda T, Uchihara JN, Taira N, Ohshiro K, Senba M, Tanaka Y, Ohshima K, Saya H, Tokino T, Mori N
  • Journal Title: Int J Cancer Vol.124, No.11 Pages: 2607-2615
  • Peer Reviewed
• [Journal Article] Cancer epigenomics : Implications of DNA methylation in personalized cancer therapy. (2009)
  • Author(s): Toyota M, Suzuki H, Yamashita T, Hirata K, Imai K, Tokino T, Shinomura Y
  • Journal Title: Cancer Sci Vol.100, No.5 Pages: 787-791
  • Peer Reviewed
• [Journal Article] Epigenetic inactivation of RASSF2 in oral squamous cell carcinoma. (2008)
  • Author(s): Imai T, Toyota M, Suzuki H, Akino K, Ogi K, Sogabe Y, Kashima L, Maruyama R, Nojima M, Mita H, Sasaki Y, Itoh F, Imai K, Shinomura Y, Hiratsuka H, Tokino T
  • Journal Title: Cancer Sci Vol.99, No.5 Pages: 958-966
  • Peer Reviewed
• [Journal Article] Epigenetic inactivation of SFRP genes in oral squamous cell carcinoma. (2008)
  • Author(s): Sogabe Y, Suzuki H, Toyata M, Ogi K, Imai T, Nojima M, Sasaki Y, Hiratsuka H, Tokino T
  • Journal Title: Int J Oncology Vol.32, No.6 Pages: 1253-1261
  • Peer Reviewed
• [Journal Article] Epigenetic silencing of microRNA-34b/c and BTG4 is associated with CpG island methylation in colorectal cancer. (2008)
  • Author(s): Toyota M, Suzuki H, Sasaki Y, Maruyama R, Imai K, Shinomura Y, Tokino T.
  • Journal Title: Cancer Res Vol.68, No.11 Pages: 4124-4132
  • Peer Reviewed
• [Journal Article] The histone deacetylase inhibitor FK228 enhances adenovirus-mediated p53 family gene therapy in cancer models. (2008)
  • Author(s): Sasaki Y, Negishi H, Idogawa M, Suzuki H, Mita H, Toyota M, Shinomura Y, Imai K, Tokino T
  • Journal Title: Mol Cancer Ther Vol.7, No.4 Pages: 779-787
  • Peer Reviewed
• [Journal Article] Identification of flotillin-2, a major protein on lipid rafts, as a novel target of the p53 family members. (2008)
  • Author(s): Sasaki Y, Oshima Y, Koyama R, Maruyama R, Akashi H, Mita H, Toyota M, Shinomura Y, Imai K, Tokino T
  • Journal Title: Mol Cancer Res Vol.6, No.3 Pages: 395-406
  • Peer Reviewed
• [Journal Article] Frequent epigenetic inactivation of Wnt antagonist genes in breast cancer. (2008)
  • Author(s): Suzuki H, Toyota M, Caraway H, Gabrielson E, Ohmura T, Fujikane T, Nishikawa N, Sogabe Y, Nojima M, Sonoda T, Mori M, Hirata K, Imai K, Shinomura Y, Baylin SB, Tokino T
  • Journal Title: Br J Cancer Vol.98, No.6 Pages: 1147-1156
  • Peer Reviewed
• [Journal Article] Cytoplasmic RASSF2A is a pro-appoptotic mediator whose expression is epigenetically silenced in gastric cancer. (2008)
  • Author(s): Maruyama R, Akino K, Toyota M, Suzuki H, Imai T, Ohe-Toyota M, Yamamoto E, Nojima M, Fujikane, Sasaki Y, Yamashita T, Watanabe Y, Hiratsuka H, Hirata K, Itoh F, Imai K, Shinomura Y, Tokino T
  • Journal Title: Carcinogenesis Vol.29, No.7 Pages: 1312-1318
  • Peer Reviewed
• [Journal Article] Arequirment for DICER to maintain full promoter CpG island hypermethylation in human cancer cells. (2008)
  • Author(s): Ting AH, Suzuki H, Cope L, Schuebel KE, Lee BH, Toyota M, Imai K, Shinomura Y, Tokino T, Baylin SB
  • Journal Title: Cancer Res Vol.68, No.8 Pages: 2570-2575
  • Peer Reviewed
• [Journal Article] DNA hypermethylation causes incomplete systhesis of carbohydrate determinants in gastrointestinal cancer. (2008)
  • Author(s): Kawamura YI, Toyota M, Kawashima R, Hagiwara T, Suzuki H, Imai K, Shinomura Y, Tokino T, Kannagi R, Dohi T
  • Journal Title: Gastroenterology Vol.135, No.1 Pages: 142-151
  • Peer Reviewed
• [Journal Article] DNA methylation and cancer pathways in gastrointestinal tumors. (2008)
  • Author(s): Suzuki H, Tokino T, Shinomura Y,Imai K, Toyota M
  • Journal Title: Pharmacogenomics Vol.9, No.12 Pages: 1917-1928
  • Peer Reviewed
• [Journal Article] LINE-1 hypomethylation is associated with increased CpG island methylation in Helicobactor pylori-related enlarged-fold gastritis. (2008)
  • Author(s): Yamamoto E, Toyota M, Suzuki H, Kondo Y, Sanomura T, Murayama Y, Ohe-Toyota M, Muruyama R, Nojima M, Ashisa M, Fujii K, Sasaki Y, Hayashi N, Mori M, Imai K, Tokino T, Shinomura Y
  • Journal Title: Cancer Epidemiol Biomarkers Prev Vol.17, No.10 Pages: 2555-2564
  • Peer Reviewed
• [Journal Article] Frequent epigenetic inactivation of SFRP genes in hepatocellular carcinoma. (2008)
  • Author(s): Takagi H, Sasaki S, Suzuki H, Toyota M, Maruyama R, Nojima M, Yamamoto H, Omata M, Tokino T, Imai K, Shinomura Y
  • Journal Title: J Gastroenterol Vol.43, No.5 Pages: 378-389
  • Peer Reviewed
• [Journal Article] In vitro transformation of adult rat hepatic progenitor cells into pancreatic endocrine hormone-producing cells. (2008)
  • Author(s): Kawasaki H, Mizuguchi T, Kikkawa Y, Oshima H, Sasaki Y, Tokino T, Kokai Y, Miyazaki J, Katsuramaki T, Mitaka T, Hirata K
  • Journal Title: J Hepatobiliary Pancreat Surg Vol.15, No.3 Pages: 310-317
  • Peer Reviewed
• [Presentation] CHFR, a potential tumor suppressor, downregulates interleukin-8 via inhibition of NF-kB. (2010)
  • Author(s): Tokino T, Kashima L, Toyota M, Sasaki Y.
  • Organizer: 101st American Association of Cancer Research (AACR) annual meeting.
  • Place of Presentation: Washington D.C
  • Year and Date: 20100400
• [Presentation] A single adenovirus expressing p53 protein and p21-targeting artificial microRNAs improves therapeutic outcome in cancer. (2010)
  • Author(s): Tokino T, Idogawa M, Sasaki Y.
  • Organizer: 8th AACR-JCA Joint International Conference.
  • Place of Presentation: Waikoloa, Hawaii
  • Year and Date: 20100200
• [Presentation] Identification of flotillin-2, a major protein on lipid rafts, as a novel target of the p53 family members. (2008)
  • Author(s): Tokino T, Sasaki Y.
  • Organizer: American Association of Cancer Research (AACR) Annual meeting.
  • Place of Presentation: San Diego, USA
  • Year and Date: 20080400
• [Remarks] ホームページ等
  • URL: http://web.sapmed.ac.jp/canmol/