Spatio-temporal analysis of cell membrane molecules involved in the enhancement of malignant properties by glycosphingolipids

• Project/Area Number: 23590371
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Pathological medical chemistry
• Research Institution: Chubu University
• Principal Investigator: FURUKAWA Keiko 中部大学, 生命健康科学部, 教授 (50260732)
• Co-Investigator(Renkei-kenkyūsha): SUZUKI Kenichi 京都大学, 物質−細胞統合システム拠点, 准教授 (50423059)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2011: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: スフィンゴ糖脂質 / メラノーマ / EMARS反応 / １分子観察 / HGF刺激 / 接着刺激 / convergence / 糖鎖合成酵素遺伝子 / 癌 / GD3 / Neogenin / 脂質ラフト / シグナル伝達

Research Abstract

We analyzed effects of GD3 expression on the cell signals triggered by hepatocyte growth factor (HGF)/Met interaction and by adhesion to collagen type I. The simultaneous treatment with both of them resulted in markedly increased activation of Akt and Erk in GD3+ cells. These results suggested that GD3 plays a crucial role in the convergence of PI3K-Akt and Ras-Erk signals, leading to the synergistic effects of those signals on malignant properties of melanomas. To search molecules existing in the vicinity of GD3 in living cells, enzyme-mediated activation of radical sources (EMARS) combined with mass spectrometry was performed. Neogenin and Integrin were identified to be interacting molecules with GD3 on lipid rafts of the cell membrane. We also tried spatio-temporal analysis of cell membrane molecules by single molecule observation, showing that glycosphingolipid GM1 specifically forms homodimer on the cell surface. Single molecule observation of GD3 on the membrane is now on-going.

Research Products

• [Journal Article] Ganglioside deficiellcy causes inflammation and neurodegeneration via the activation of complement system in the spinal cord. (2014)
  • Author(s): Yuhsuke Ohmi, Yuki Ohkawa, et al.
  • Journal Title: Journal of Neuroinflammation Volume: 61(印刷中) Issue: 1 Pages: 61-61
  • DOI: 10.1186/1742-2094-11-61
  • Peer Reviewed / Open Access
• [Journal Article] UVB-irradiated keratinocytes induce melanoma-associated ganglioside GD3 synthase gene in melanocytes via secretion of tumor necrosis factor α and interleukin 6. (2014)
  • Author(s): Miyata M.
  • Journal Title: Biochem Biophys Res Commun. Volume: 445 Issue: 2 Pages: 504-10
  • DOI: 10.1016/j.bbrc.2014.02.038
  • Peer Reviewed / Open Access
• [Journal Article] Gagnlioside GD3 induces convergence and synergism of adhesion and hepatocyte growth factor/Met signals in melanomas. (2014)
  • Author(s): Keiko Furukawa, Yuki Ohkawa, et al.
  • Journal Title: Cancer Science Volume: 105 Issue: 1 Pages: 52-63
  • DOI: 10.1111/cas.12310
  • Peer Reviewed
• [Journal Article] Neuronal expression of GalNAc transferase is sufficient to prevent the age-related neurodegenerative phenotype of complex ganglioside-deficient mice. (2014)
  • Author(s): Yao D.
  • Journal Title: J Neurosci. Volume: 34 Pages: 880-91
  • Peer Reviewed
• [Journal Article] Binding of a sialic acid-recognizing lectin Siglec-9 modulates adhesion dynamics of cancer cells via calpain-mediated protein degradation. (2013)
  • Author(s): Sabit I.
  • Journal Title: J Biol Chem. Volume: 288 Issue: 49 Pages: 35417-27
  • DOI: 10.1074/jbc.m113.513192
  • Peer Reviewed
• [Journal Article] β4GalT6 is involved in the synthesis of lactosylceramide with less intensity than β4GalT5 (2013)
  • Author(s): Tokuda N, Numata S, Li X, Nomura T, Takizawa M, Kondo Y, Yamashita Y, Hashimoto N, Kiyono T, Urano T, Furukawa K, Furukawa K
  • Journal Title: Glycobiology Volume: 23 Issue: 10 Pages: 1175-1183
  • DOI: 10.1093/glycob/cwt054
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Trimeric Tn antigen on Syndecan-1 binds to integrins and enhances cancer metastasis via dynamic changes of membrane microdomains. (2013)
  • Author(s): Matsumoto Y, Zhang Q, Akita K, Nakada H, Hamamura K, Tsuchida A, Okajima T, Furukawa K, Urano T, Furukawa K
  • Journal Title: J. Biol. Chem. Volume: 288 Issue: 33 Pages: 24264-24276
  • DOI: 10.1074/jbc.m113.455006
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] TLR4-MD-2 complex is negatively regulated by an endogenous ligand, globotetraosylceramide in vascular endothelial cells. (2013)
  • Author(s): Kondo, Y., Tokuda, N., Nishitani, C., Ohto, U., Akashi-TakKondo, Y., Tokuda, N., Nishitani, C., Ohto, U., Akashi-Takamura, S., Ito, Y., Uchikawa, M., Kuroki, Y., Miyake, K., Zhang, Q., Furukawa, K., Furukawa, K
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 110 Issue: 12 Pages: 4714-4719
  • DOI: 10.1073/pnas.1218508110
  • Peer Reviewed
• [Journal Article] Disialyl gangliosides enhance tumor phenotypes with differential modalities. (2012)
  • Author(s): Koichi Furukawa
  • Journal Title: Glycqconjugate journal Volume: 29 Issue: 8-9 Pages: 579-584
  • DOI: 10.1007/s10719-012-9423-0
  • Peer Reviewed
• [Journal Article] Proteomic analysis of ganglioside-associated membrane molecules: Substantial basis for molecular clustering. (2012)
  • Author(s): Hashimoto, N., Hamamura, K., Kotani, N., Furukawa, K., Kaneko, K., Honke, K., Furukawa, K.
  • Journal Title: Proteomics Volume: 12 Issue: 21 Pages: 3154-3163
  • DOI: 10.1002/pmic.201200279
  • Peer Reviewed
• [Journal Article] Enhancement of malignant properties of human osteosarcoma cells with disialyl gangliosides GD2/GD3. (2012)
  • Author(s): Shibuya H.
  • Journal Title: Cancer Sci. Volume: 103 Issue: 9 Pages: 1656-1664
  • DOI: 10.1111/j.1349-7006.2012.02344.x
  • Peer Reviewed
• [Journal Article] Fine tuning of cell signals by glycosylation (2012)
  • Author(s): Furukawa K, Ohkawa Y, Yamauchi Y, Hamamura K, Ohmi Y, Furukawa K.
  • Journal Title: J Biochem. Volume: 151 Issue: 6 Pages: 573-578
  • DOI: 10.1093/jb/mvs043
  • NAID: 10031166098
  • Peer Reviewed
• [Journal Article] Essential roles of gangliosides in the formation and maintenance of membrane microdomains in brain tissues. (2012)
  • Author(s): Yuhsuke Ohmi
  • Journal Title: Neurochemical Research Volume: 37 Issue: 6 Pages: 1185-1191
  • DOI: 10.1007/s11064-012-0764-7
  • Peer Reviewed
• [Journal Article] Clostridium perfringens Alpha-toxin Recognizes the GM1a/TrkA Complex. (2012)
  • Author(s): Oda M.
  • Journal Title: J. Biol. Chem. Volume: 287 Pages: 33070-33079
  • Peer Reviewed
• [Journal Article] Lewis y antigen is expressed in oral squamous cell carcinoma cell lines and tissues, but disappears in the invasive regions leading to the enhanced malignant properties irrespective of sialyl-Lewis x. (2012)
  • Author(s): Hotta H.
  • Journal Title: Glycoconj J. Volume: -
  • Peer Reviewed
• [Journal Article] pp-GalNAc-T13 induces high metastatic potential of murine Lewis lung cancer by generating trimeric Tn antigen. (2012)
  • Author(s): Matsumoto Y.
  • Journal Title: Biochem Biophys Res Commun. Volume: 419 Issue: 1 Pages: 7-13
  • DOI: 10.1016/j.bbrc.2012.01.086
  • Peer Reviewed
• [Journal Article] Functional activation of Src family kinase Yes is essential for the enhanced malignant properties of human melanoma cells expressing ganglioside GD3 (2011)
  • Author(s): Hamamura, K., Tsuji, M., Hotta, H., Ohkawa, Y., Takahashi, M., Shibuya, H., Nakashima, H., Yamauchi, Y., Hashimoto, N., Hattori, H., Ueda, M., Furukawa, K., Furukawa. K
  • Journal Title: J.Biol.Chem. Volume: 286 Issue: 21 Pages: 18526-37
  • DOI: 10.1074/jbc.m110.164798
  • Peer Reviewed
• [Journal Article] Wisp2/CCN5 up-regulated in the central nervous system of GM3-only mice facilitates neurite formation in Neuro2a cells via integrin-Akt signaling (2011)
  • Author(s): Ohkawa, Y., Ohmi, Y, Tajima, O., Yamauchi, Y, Furukawa, K., Furukawa' K
  • Journal Title: Biochem.Biophys.Res.Commun. Volume: 411 Issue: 3 Pages: 483-489
  • DOI: 10.1016/j.bbrc.2011.06.118
  • Peer Reviewed
• [Journal Article] Positive feedback loop between PI3K-Akt-mTORC1 signaling and the lipogenic pathway boosts Akt signaling : induction of the lipgenic pathway by a melanoma antigen (2011)
  • Author(s): Yamauchi Y, Furukawa K, Hamamura K, Furukawa K.
  • Journal Title: Cancer Res. Volume: 71 Issue: 14 Pages: 4989-4997
  • DOI: 10.1158/0008-5472.can-10-4108
  • Peer Reviewed
• [Journal Article] Membrane sialidase NEU3 is highly expressed in human melanoma cells promoting cell growth with minimal changes in the composition of gangliosides (2011)
  • Author(s): Miyata M., Kambe M., Tajima O., Moriya S., Sawaki H., Hotta H., Kondo Y., Narimatsu H., Miyagi T., Furukawa K., Furukawa K.
  • Journal Title: Cancer Sci Volume: 102 Issue: 12 Pages: 2139-2149
  • DOI: 10.1111/j.1349-7006.2011.02086.x
  • NAID: 10030245717
  • Peer Reviewed
• [Journal Article] Molecular Network Associated with MITF in Skin Melanoma Development and Progression. (2011)
  • Author(s): Ya manoshita O Kato M
  • Journal Title: J Skin Cancer. Volume: 2011 Pages: 730170-730170
  • DOI: 10.1155/2011/730170
  • Peer Reviewed
• [Journal Article] Regulatory mechanisms of nervous systems with glycosphingolipids. (2011)
  • Author(s): Furukawa K.
  • Journal Title: Neurochem Res. Volume: 36 Issue: 9 Pages: 1578-1586
  • DOI: 10.1007/s11064-011-0494-2
  • Peer Reviewed
• [Journal Article] Efficient generation of useful monoclonal antibodies reactive with globotriaosylceramide using knockout mice lacking Gb3/CD77 synthase. (2011)
  • Author(s): Kondo Y.
  • Journal Title: Glycoconj J. Volume: 28 Issue: 6 Pages: 371-384
  • DOI: 10.1007/s10719-011-9335-4
  • Peer Reviewed
• [Presentation] シグレック9の結合はカルパインによりタンパク質分解に基づいて癌細胞の接着動態を調節する (2013)
  • Author(s): 伊力哈木江
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜
• [Presentation] GD3発現ヒトメラノーマ細胞におけるNeogeninの役割 (2013)
  • Author(s): 金子慶
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜
• [Presentation] DGFBによって発現誘導されるガングリオシドGD3はグリオーマ細胞の悪性形質を増強する (2013)
  • Author(s): 大川祐樹
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜
• [Presentation] 複合糖質と糖鎖認識分子との相互作用の時空間的ダイナミクス (2013)
  • Author(s): 古川鋼一
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] アストロサイトにおけるガンクリオシド糖鎖の機能 (2013)
  • Author(s): 大海雄介
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] ガングリオシドによるamyloidβprecursor protein(APP)の切断パターンの制御及びAPP切断断片が細胞の健常性に与える影響 (2013)
  • Author(s): 山口世堯
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] 脂肪細胞のレプチン分泌と褐色脂肪細胞におけるb細胞列ガングリオシドの調節機構 (2013)
  • Author(s): 姫しゅうてい
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] 糖鎖変異による脂質吸収障害の分子メカニズム (2013)
  • Author(s): 田島織絵
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] TNFα及び紫外線によるメラノサイトとメラノーマのGD3合成酵素遺伝子の発現誘導 (2013)
  • Author(s): 古川圭子
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] 複合糖質と糖鎖認識分子との相互作用の時空間的ダイナミクス (2013)
  • Author(s): 古川 鋼一
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] GD3発現メラノーマの悪性形質におけるNeogeninの機能 (2013)
  • Author(s): 金子 慶
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] アストロサイトにおけるガンクリオシド糖鎖の機能 (2013)
  • Author(s): 大海 雄介
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] ガングリオシドによるamyloid  precursor protein (APP) の切断パターンの制御及びAPP切断断片が細胞の健常性に与える影響 (2013)
  • Author(s): 山口 世堯
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] 脂肪細胞のレプチン分泌と褐色脂肪細胞におけるb細胞列ガングリオシドの調節機構 (2013)
  • Author(s): 姫 しゅうてい
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] 糖鎖変異による脂質吸収障害の分子メカニズム (2013)
  • Author(s): 田島 織絵
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] 腎癌細胞株における分岐型ジシアリル糖鎖抗原の役割 (2013)
  • Author(s): 土田 明子
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] Biding of Siglec-9 to the counter-receptors on cancer cells regulates adhesion dynamics via protease-mediated degradation of focal adhesion kinase (FAK) (2013)
  • Author(s): Yilihamujiang Sabit
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] TNFα及び紫外線によるメラノサイトとメラノーマのGD3合成酵素遺伝子の発現誘導 (2013)
  • Author(s): 古川 圭子
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] シグレック9の結合はカルパインによりタンパク質分解に基づいて癌細胞の接着動態を調節する (2013)
  • Author(s): 伊力哈木江
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] GD3発現ヒトメラノーマ細胞におけるNeogeninの役割 (2013)
  • Author(s): 金子 慶
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] TNFαによるメラノサイトおよびメラノーマにおけるGD3合成酵素遺伝子の発現誘導 (2013)
  • Author(s): 宮田 麻衣子
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] PDGFBによって発現誘導されるガングリオシドGD3はグリオーマ細胞の悪性形質を増強する (2013)
  • Author(s): 大川 祐樹
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] ヒト口腔扁平上皮癌に関して、Lewis y 抗原の発現と進行期における消失 (2013)
  • Author(s): 堀田 宏司
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜、横浜市
• [Presentation] GD3発現細胞での接着とHGF刺激によるAktの相乗的活性化とアポトーシス抑制 (2012)
  • Author(s): 古川圭子
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡国際会議場
• [Presentation] 腸管上皮細胞におけるスフィンゴ糖脂質の役割 (2012)
  • Author(s): 田島織絵
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡国際会議場
• [Presentation] Expression profiling of glycosyltransferase genes in human melanocytes and melanoma cells (2012)
  • Author(s): Miyata Maiko
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: ロイトン札幌
• [Presentation] 糖鎖変異による小腸機能障害の分子メカニズム (2012)
  • Author(s): 田島織絵
  • Organizer: 第31回日本糖質学会年会
  • Place of Presentation: 鹿児島市民文化ホール
• [Presentation] ヒト メラノサイトおよびメラノーマ細胞における糖転移酵素遺伝子発現プロファイリング (2012)
  • Author(s): 宮田麻衣子
  • Organizer: 第76回日本生化学会中部支部例会
  • Place of Presentation: 愛知県 岡崎市
• [Presentation] 糖鎖変異による小腸機能障害の分子メカニズム (2012)
  • Author(s): 田島織絵
  • Organizer: 第31回日本糖質学会年会
  • Place of Presentation: 鹿児島市
• [Presentation] Expression profiling of glycosyltransferase genes in human melanocytes and melanoma cells (2012)
  • Author(s): Miyata Maiko
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: 札幌市
• [Presentation] GD3発現細胞での接着とHGF刺激によるAktの相乗的活性化とアポトーシス抑制 (2012)
  • Author(s): 古川圭子
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡市
• [Presentation] 腸管上皮細胞におけるスフィンゴ糖脂質の役割 (2012)
  • Author(s): 田島織絵
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡市
• [Presentation] Egr-1によるCD109遺伝子の発現制御機構の解析 (2012)
  • Author(s): 祖父江沙矢加
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡市
• [Presentation] メラノーマ特異的酸性スフィンゴ糖脂質によるc-Met機能の増強メカニズム (2011)
  • Author(s): 古川圭子
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館
• [Presentation] Gut abnormalities during postnatal development in GM3-only mice (2011)
  • Author(s): 田島織絵
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館
• [Presentation] Convergence and enhancement of the growth and adhesion signals by ganglioside GD3in human melanomas (2011)
  • Author(s): Furukawa Keiko
  • Organizer: 21st International Symposium on Glycoconjugates
  • Place of Presentation: Austria
• [Presentation] Molecular mechanisms for the growth disorders in GM3-only mice (2011)
  • Author(s): Tajima Orie
  • Organizer: 21st International Symposium on Glycoconjugates
  • Place of Presentation: Austria
• [Presentation] メラノーマ特異的酸性スフィンゴ糖脂質によるc-Met機能の増強メカニズム. (2011)
  • Author(s): 古川圭子
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館
• [Presentation] Gut abnormalities during postnatal development in GM3-only mice. (2011)
  • Author(s): 田島織絵
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館
• [Presentation] Convergence and enhancement of the growth and adhesion signals by ganglioside GD3 in human melanomas. (2011)
  • Author(s): Furukawa K.
  • Organizer: 21st International Symposium on Glycoconjugates
  • Place of Presentation: ウィーン大学、Austria
• [Presentation] Molecular mechanisms for the growth disorders in GM3-only mice. (2011)
  • Author(s): Tajima O.
  • Organizer: 21st International Symposium on Glycoconjugates
  • Place of Presentation: ウィーン大学、Austria
• [Book] Lipid Rafts (Editor:Dan Sillence), The Role of Glycosphingolipids in Lipid Rafts (2013)
  • Author(s): Koichi Furukawa, Yusuke Ohmi, Yuji Kondo, Yuki Ohkawa, Noboru Hashimoto, Orie Tajima, Keiko Furukawa.
  • Publisher: Nova Science Publishers Inc.
• [Book] シグナル伝達研究最前線2012糖鎖修飾によるシグナル伝達の制御((増刊)Vol.30) (2012)
  • Author(s): 古川鋼一, 浜村和紀, 大川祐樹, 大海雄介, 古川圭子, 羊土社
  • Publisher: 実験医学
• [Book] シグナル伝達 研究最前線2012：糖鎖修飾によるシグナル伝達の制御 (2012)
  • Author(s): 古川 鋼一
  • Publisher: 羊土社、実験医学（増刊）Vol. 30