Evaluation of GPR48 as a novel molecular target which is involved in the tumor progression

• Project/Area Number: 19590064
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Hamamatsu University School of Medicine
• Principal Investigator: KITAGAWA Kyoko Hamamatsu University School of Medicine, 医学部, 助教 (20299605)
• Co-Investigator(Kenkyū-buntansha): KITAGAWA Masatoshi 浜松医科大学, 医学部, 教授 (50294971)
• Co-Investigator(Renkei-kenkyūsha): KITAGAWA Masatoshi 浜松医科大学, 医学部, 教授 (50294971)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2008: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2007: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: 分子生物学 / GPR / 癌

Research Abstract

細胞浸潤能変化
invasion assayによりGPR48発現量に正相関した浸潤能の亢進が細胞レベルで認められていたため、そのメカニズムを探るため、細胞運動能の変化をscratch assayで調べた。使用した恒常発現株のGPR48発現量は株間で様々であったが、運動能との相関性は見出せなかった。さらに浸潤能亢進レベルも対照群と有為差が認められなくなった。このため、細胞外マトリックス分解能などの測定も計画していたが中止とした。
発癌促進効果
GPR48遺伝子を導入した培養細胞でcolony formation assayとfocus assayを行い、GPR48発現亢進による発癌性への寄与を検討したが、発癌促進効果は認められなかった。しかし細胞レベルでの研究から、GPR48は癌の悪性度を促進する作用を示した。
GPR48トランスジェニックマウスの解析
全身性発現型トランスジェニックマウスの作成に成功し、外来性GPR48発現臓器の異なる3ラインを得た。個体レベルでの機能解析は、ノックアウトマウスにおいて発育不全や腎臓形成不全が認められているが、全身性発現型GPR48トランスジェニックマウスでは発育程度、腎形成の異常は出現せず、長期通常飼育下で癌その他の疾患の発生の傾向は認められなかった。
我々はこれまでの他の研究プロジェクトにより、p27のユビキチンプロテアソームシステムによる分解の基質認識タンパクであるSkp2ホモノックアウトマウスは、尿管結紮による腎不全モデル(UUO マウス)を作成すると腎障害レベルがヘテロまたはホモ野生型と比較して著明に軽減されていることを見出していた。このような表現系の発現に、Skp2によるp27分解の抑制、さらにGPR48発現量の上昇が関与している可能性を考え、GPR48トランスジェニックマウスでUUOを作成して組織学的に解析した。その結果、腎障害の程度はコントロールUUOマウスと同等であり、Skp2ノックアウトマウスで見られた腎の特徴はGPR48を介してはいないと考えられた。これらのマウスを用いた結果から、GPR48はp27の下流因子の一つではあるものの、腎障害および発癌の過程においてGPR48がp27下流の主経路に関与している可能性は低いと思われた。

Research Products

• [Journal Article] High expression of Pirh2, an E3 ligase for p27, is associated with low expression of p27 and poor prognosis in head and neck cancers (2009)
  • Author(s): Shimada, M., et al.
  • Journal Title: Cancer Sci. (In press)
  • Peer Reviewed
• [Journal Article] Effects of 5-week ethanol feeding on the liver of aldehyde dehydrogenase 2 knockout mice (2008)
  • Author(s): Matsumoto, A, Kawamoto, T, Mutoh,F, Isse, T, Oyama, T, Kitagawa, K, Nakayama, K.-i.and Ichiba, M.
  • Journal Title: Pharmacogenet Genomics 18(10) Pages: 847-52
• [Journal Article] Increased frequencies of micronucleated reticulocytes and T-cell receptor mutation in Aldh2 knockout mice exposed to acetaldehyde (2008)
  • Author(s): Kunugita, N, Isse, T, Oyama, T, Kitagawa, K, Ogawa, M, Yamaguchi,T, Kinaga, T,and Kawamoto, T.
  • Journal Title: J.Toxicol. Sci 33(1) Pages: 31-6
  • NAID: 110006596029
• [Journal Article] TSU68 prevents liver metastasis of colon cancer xenografts by modulating the pre-metastatic niche (2008)
  • Author(s): Yamamoto, M, Kikuchi, H, Ohta, M, Kawabata, T, Hiramatsu, Y, Kondo, K, Baba, M, Kamiya, K, Tanaka, T, Kitagawa and M. Konno H.
  • Journal Title: Cancer Res 68 Pages: 9754-9762
• [Journal Article] Intrarenal RAS activity and urinary angiotensinogen excretion in anti-thymocyte serum nephritis rats (2008)
  • Author(s): Ohashi, N, Yamamoto, T, Huang, Y, Misaki, T, Fukasawa, H, Suzuki, H, Togawa, A, Suzuki, S, Fujigaki, Y, Nakagawa, T, Nakamura, Y, Suzuki, F, Kitagawa, M, Hishida, A.
  • Journal Title: Am J.Physiol Renal Physiol 295 Pages: 1512-1518
• [Journal Article] Novel insights into FGD3, a putative GEF for Cdc42, that undergoes SCF^<FWD1/b-TrCP>-mediated proteasomal degradation analogous to that of its homologue FGD1 but regulates cell morphology and motility differently from FGD1 (2008)
  • Author(s): Hayakawa, M, Matsushima, M, Hagiwara, H, Oshima, T, Fujino, T, Ando, K, Kikugawa, K, Tanaka, H, Miyazawa,K.and Kitagawa, M.
  • Journal Title: Genes to Cells 13 Pages: 329-442
• [Journal Article] Pirh2 interacts with and ubiquitylates signal recognition particle receptor beta subunit (2008)
  • Author(s): Abe, K, Hattori, T, Isobe, T, Kitagawa, K, Oda, T, Uchida, C.and Kitagawa, M.
  • Journal Title: Biomed Res 29 Pages: 53-60
  • NAID: 130004470702
• [Journal Article] Stress via p53 pathway causes apoptosis by mitochondrial Noxa upregulation in doxorubicin-treated neuroblastoma cells (2008)
  • Author(s): Kurata, K, Yanagisawa, R., Ohira, M. Kitagawa, M, Nakagawara, A, Kamijo, T.
  • Journal Title: Oncogene 27 Pages: 741-754
• [Journal Article] Regulation of TGF-beta signaling by Smads and its roles in tissue fibrosis (2008)
  • Author(s): Fukasawa, H. Yamamato, T. Kitagawa, M. and Hishida A.
  • Journal Title: Current signal transduction therapy 3 Pages: 1-6
• [Journal Article] Effects of 5-week ethanol feeding on the liver of aldehy dedehydrogenase 2 knockout mice. (2008)
  • Author(s): Matsumoto, A, et al.
  • Journal Title: Pharmacogenet Genomics 18 Pages: 847-852
  • Peer Reviewed
• [Journal Article] Pirh2 interacts with and ubiquitylates signal recognition particle receptor beta subunit (2008)
  • Author(s): Abe, K., et. al.
  • Journal Title: Biomed Res. 29 Pages: 53-60
  • Peer Reviewed
• [Journal Article] Increased frequencies of micronucleated reticulocytes and T-cell receptor mutation in Aldh2 knockout mice exposed to acetaldehyde. (2008)
  • Author(s): Kunugita, N., et. al.
  • Journal Title: J Toxicol Sci. 33 Pages: 31-36
  • Peer Reviewed
• [Journal Article] Reproductive Toxicity of Ethylene Glycol Monoethyl Ether in Aldh2 Knockout Mice (2007)
  • Author(s): Wang, R.S, Ohtani, K, Suda, M, Kitagawa, K, Nakayama, K.-I, Kawamoto, T,and Nakajima, T.
  • Journal Title: Industrial Health 45(4) Pages: 574-578
  • NAID: 130004828493
• [Journal Article] Increased formation of hepaticN2-ethylidene-2'-deoxyguanosine DNA adducts in aldehyde dehydrogenase 2 knockout mice treated with ethanol (2007)
  • Author(s): Matsuda, T, Matsumoto, A, Uchida, M, Kanaly, R., Misaki, K, Shibutani, S, Kawamoto, T, Kitagawa, K, Nakayama, K.-I, Tomokuni, K,and Ichiba, M.
  • Journal Title: Carcinogenesis 28(11) Pages: 2363-2366
• [Journal Article] Pirh2 Promotes Ubiquitin-Dependent Degradation of the CDK Inhibitor p27Kip1 (2007)
  • Author(s): Hattori, T, Isobe, T, Abe, K, Kikuchi, H, Kitagawa, K, Oda, T, Uchida, C,and Kitagawa, M.
  • Journal Title: Cancer Research 67 Pages: 10789-10795
• [Journal Article] Critical and functional regulation of CHOP(C/EBP homologous protein)through the N-terminal portion (2007)
  • Author(s): Ohoka, N, Hattori, T, Kitagawa, M, Onozaki, K,and Hayashi H.
  • Journal Title: J Biol Chem 282 Pages: 35687-35694
• [Journal Article] Contribution of Extracellular Signal-Regulated Kinases to the IL-1-induced Growth Inhibition of Human Melanoma Cells A375 (2007)
  • Author(s): Arakawa, T, Yamamura, T, Hattori, T, Hayashi H, Mori, A, Yoshida, A, Uchida, C, Kitagawa, M, and Onozaki, K.
  • Journal Title: International Immunopharmacology 8 Pages: 80-89
• [Journal Article] Cys-881 is essential for the trafficking and secretion of truncated mutant ceruloplasmin in aceruloplasminemia (2007)
  • Author(s): Kono, S, Suzuki, H, Oda, T, Takahashi, Y, Shirakawa, K, Takahashi, Y, Kitagawa, M,and Miyajima, H.
  • Journal Title: J. Hepatology 47 Pages: 844-850
• [Journal Article] Effect of loss of heterozygosity of the c-kit gene on prognosis after hepatectomy for metastatic liver gastrointestinal stromal tumors (2007)
  • Author(s): Kikuchi, H, Yamamoto, M, Hiramatsu, Y, Baba, M, Ohta, M, Kamiya, K, Tanaka, T, Suzuki, S, Sugimura, H, Kitagawa, M, Kanai, T, Kitayama, Y, Kanda, T, Nishikura, K,and Konno, H.
  • Journal Title: Cancer Sci 98 Pages: 1734-1739
  • NAID: 10020119012
• [Journal Article] ARA54 is involved in transcriptional regulation of the cyclin D1 gene in human cancer cells (2007)
  • Author(s): Kikuchi, H, Uchida, C, Hattori, T, Isobe, T, Hiramatsu, Y, Kitagawa, K, Oda, T, Konno, H,and Kitagawa, M.
  • Journal Title: Carcinogenesis 28 Pages: 1752-1758
• [Journal Article] Renal damage in obstructive nephropathy is decreased in Skp2-deficient mice (2007)
  • Author(s): Suzuki, S, Fukasawa, H, Kitagawa, K, Uchida, C, Hattori, T, Isobe, T, Oda, T, Misaki, T, Ohashi, N, Nakayama, K, Nakayama, K.I., Hishida, A., Yamamoto, T,and Kitagawa, M.
  • Journal Title: Am.J.Pathol 171 Pages: 473-483
• [Journal Article] Phosphorylation of pRB at Ser612 by Chk1/2 leads to a complex between pRB and E2F-1 after DNA damage (2007)
  • Author(s): Inoue, Y, Kitagawa, M,and Taya, Y.
  • Journal Title: EMBO J 26 Pages: 2083-2093
• [Journal Article] Genetically engineered neural stem cells migrate and suppress glioma cell growth at distant intracranial sites (2007)
  • Author(s): Li, S, Gao, Y, Tokuyama, T, Yamamoto, J, Yokota, N, Yamamoto, S, Terakawa, S, Kitagawa, M,and Namba, H.
  • Journal Title: Cancer Letters 251(2) Pages: 220-227
• [Journal Article] Increased formation of hepatic N2-ethylidene-2'-deoxyguanosine DNA adducts in aldehy dedehydrogenase 2-knockout mice treated with ethanol (2007)
  • Author(s): Matsuda, T., et. al.
  • Journal Title: Carcinogenesis 28 Pages: 2363-2366
  • Peer Reviewed
• [Journal Article] Pirh2 Promotes Ubiquitin・Dependent Degradation of the CDK Inhibitor p27Kip1 (2007)
  • Author(s): Hattori, T., et. al.
  • Journal Title: Cancer Res. 67 Pages: 10789-10795
  • Peer Reviewed
• [Journal Article] ARA54 is involved in transcriptional regulation of the cyclin Dl gene in human cancer cells (2007)
  • Author(s): Kikuchi, H., et. al.
  • Journal Title: Carcinogensis 28 Pages: 1752-1758
  • Peer Reviewed
• [Journal Article] Renal damage in obstructive nephropathy is decreased in Skp2-deficient mice (2007)
  • Author(s): Suzuki, S., et. al.
  • Journal Title: Am.J.Pathol. 171 Pages: 473-483
  • Peer Reviewed
• [Journal Article] Reproductive toxicity of ethylene glycol monoethyl ether in Aldh2 knock out mice (2007)
  • Author(s): Wang, R.S., et. al.
  • Journal Title: Ind.Health 45 Pages: 574-578
  • Peer Reviewed
• [Journal Article] Effect of loss of heterozygosity of the c-kit gene on prognosis after hepatectomy for metastatic liver gastrointestinal stromal tumors (2007)
  • Author(s): Kikuchi, H., et. al.
  • Journal Title: Cancer Sci. 98 Pages: 1734-1739
  • Peer Reviewed
• [Journal Article] Cys-881 is essential for the trafficking and secretion of truncated mutant ceruloplasmin in aeruloplasminemia (2007)
  • Author(s): Kono, S., et. al.
  • Journal Title: J.Hepatology 47 Pages: 844-850
  • Peer Reviewed
• [Journal Article] Stress via p53 pathway causes apoptosis by mitochondrial Noxa upregul ation in doxorubicin-treated neuroblastoma cells (2007)
  • Author(s): Kurata, K., et. al.
  • Journal Title: Oncogene 27 Pages: 741-754
  • Peer Reviewed
• [Journal Article] Critical and functional regulation of CHOP(CIEBP homologous protein) through the N-terminal portion (2007)
  • Author(s): Ohoka, N, , et. al.
  • Journal Title: J Biol Chem 282 Pages: 35687-35694
  • Peer Reviewed
• [Journal Article] Contribution of Extracellular Signal-Regulated Kinases to the IL-1-induced Growth Inhibition of Human Melanoma Cells A375 (2007)
  • Author(s): Arakawa, T., et. al.
  • Journal Title: International Immunopharmacology 8 Pages: 80-89
  • Peer Reviewed
• [Presentation] GPR48による癌転移機構の解析 (2008)
  • Author(s): 磯部智康, 島田真衣, 劉寧, 北川恭子, 鈴木小由里, 小田敏明, 服部隆行, 内田千晴, 北川雅敏
  • Organizer: 第31回日本分子生物学会、第81回日本生化学会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-10
• [Presentation] タンパク質過剰発現誘導性小胞体の構造と機能 (2008)
  • Author(s): 小田敏明, 服部隆行, 磯部智康、北川恭子, 内田千晴, 横田貞記, 北川雅敏
  • Organizer: 第31回日本分子生物学会、第81回日本生化学会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-09
• [Presentation] 細胞周期制御分子p27のユビキチン依存的分解機構と病態 (2007)
  • Author(s): 北川雅敏
  • Organizer: 第30回日本分子生物学会第80回日本生化学会合同大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-12
• [Presentation] 細胞周期制御分子p27のユビキチン依存的分解機構と病態 (2007)
  • Author(s): 北川 雅敏
  • Organizer: 第30回日本分乎生物学会第80回日本生化学会合同大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-12
• [Presentation] Novel regulatory mechanisms of Cyclin D1 expression (2007)
  • Author(s): Kitagawa, M, et.al.
  • Organizer: 第66回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2007-10-05
• [Presentation] Novel regulatory mechanisms of Cyclin Dl expression (2007)
  • Author(s): Kitagawa, M., et. al.
  • Organizer: 第66回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2007-10-05
• [Presentation] 癌関連核内因子のユビキチン依存的制御機構 (2007)
  • Author(s): 北川雅敏
  • Organizer: Nuclear Signaling JAPAN 2007シンポジウム
  • Place of Presentation: 東京
  • Year and Date: 2007-07-13
• [Remarks]
  • URL: http://www2.hama-med.ac.jp/w1a/bio1/index-j.html