2003 Fiscal Year Annual Research Report

小児癌由来cDNAチップを用いた正常個体発生の遺伝子発現ネットワークの網羅的解析

Research Project

Project/Area Number	15012254
Research Institution	Chiba Cancer Center (Research Institute)
Principal Investigator	大平美紀千葉県がんセンター, 生化学研究部, 研究員 (20311384)
Co-Investigator(Kenkyū-buntansha)	中川原章千葉県がんセンター, 生化学研究部, 部長 (50117181)
Keywords	神経芽腫 / 肝芽腫 / 個体発生 / DNAチップ
Research Abstract	本研究では、発生初期の神経芽細胞や肝細胞、腎細胞の発生分化に関わる遺伝子群の同定と解析を目指し、小児癌由来遺伝子を搭載したチップの作製と、神経芽細胞が交感神経系へ分化成熟する過程で働く転写因子群およびWntシグナル等に関わる遺伝子群の遺伝子発現ネットワークの解明を目的としている。本年度は以下を行った。 1)遺伝子発現系の作製と神経芽腫由来チップを用いた網羅的発現解析アデノウイルスベクターに組み込んだMASH1、Phox2a,Phox2b遺伝子を神経芽腫細胞株に導入し、ノザン解析及びRT-PCRによる経時的な発現誘導を確認した。MYCN遺伝子については、テトラサイクリン除去によりMYCNが発現誘導される神経芽腫細胞株を用い、これまでに作製した神経芽腫由来の約5000種類の遺伝子を搭載したチップを用いてハイブリダイゼーションを行った。同定された候補遺伝子の半定量RT-PCRおよびノザン解析にてチップ解析の再現性を確認した結果、少なくとも3つのMYCN下流遺伝子の候補を同定した。さらに、神経芽腫の予後良好群と不良群の間で顕著に異なる発現を示す300以上の遺伝子群も同定した。これらは、神経堤細胞の分化成熟に何らかの関与をしている可能性が考えられた。また、これらの遺伝子から神経芽腫の腫瘍の個性と相関する遺伝子の候補をクラスタリング解析等により同定し、ランキングを行った。これらは神経芽腫の悪性度を予測する良いモデルケースになると期待された。 2)肝芽腫チップの作製と腎芽腫遺伝子ライブラリーの作製 β-cateninに変異がある肝芽腫、変異のない肝芽腫、及び非癌部の3つを出発材料として合計約12,000cDNAクローンからなる遺伝子ソースを調製し、約5,000の独立遺伝子のチップ作製を行った。また、半定量RT-PCRによる癌部と非癌部の間で発現に差のある遺伝子のスクリーニングを行い、約80の遺伝子を同定した。さらに、腎芽腫の腫瘍および非癌部から12,000クローンのcDNAを蒐集し、両端塩基配列決定を終えた。

Research Products

(13 results)

All Other

All Publications (13 results)

[Publications] Nakagawara A, Ohira M.: "Comprehensive genomics linking between neural development and cancer : Neuroblastoma as a model. In Special Issue : Neural development and cancer."Cancer Letters. (in press).
[Publications] Hamano S, Ohira M, Isogai E, Nakada K, Nakagawara A: "Identification of Novel Human Neuronal Leucine-Rich Repeat (hNLRR) Family Genes and Inverse Association of Expression of Nbla10449/hNLRR-1 and Nbla10677/hNLRR-3 with the Prognosis of Primary Neuroblastomas"Int.J Oncology. (in press).
[Publications] Ohira M, Morohashi A, Inuzuka H, Nakagawara A, et al.: "Expression profiling and characterization of 4200 genes cloned from primary neuroblastomas : identification of 305 genes differentially expressed between favorable and unfavorable subsets"Oncogene. 22. 5525-5536 (2003)
[Publications] Ohira M, Morohashi A, Nakamura Y, Nakagawara A, et al.: "Neuroblastoma oligo-capping cDNA project : toward the understanding of the genesis and biology of neuroblastoma"Cancer Letters. 197. 63-68 (2003)
[Publications] Kawamoto T, Ohira M, Hamano S, Hori T, Nakagawara A: "High expression of the ovel endothelin-converting enzyme genes, Nbla03145/ECEL1α and β, is associated with favorable prognosis in human neuroblastomas"Int J Oncology. 22. 815-822 (2003)
[Publications] Tomioka N, Kobayashi H, Kageyama H, Ohira M, Nakagawara A, et al.: "Chromosomes that show partial loss or gain in near-diploid tumors coincide with chromosomes that show whole loss or gain in near-triploid tumors : Evidence suggesting the involvement of the same genes in the tumorigenesis of high- and low-risk neuroblastomas"Genes Chromosomes Cancer. 36. 139-150 (2003)
[Publications] Nakagawara A: "Neural crest development and neuroblastoma : The genetic and biological link"Neuroscience Series, Progress in Brain Research, Elsevier Science Publisher. (in press).
[Publications] Tomizawa M, Watanabe K, Saisho H, Nakagawara A, Tagawa M.: "Down-regulated expression of the CCAAT/enhancer binding protein a and b genes in human hepatocellular carcinoma : a possible prognostic marker"Anticancer Res. 23. 351-354 (2003)
[Publications] Saito-Ohara F, Imoto I, Inoue J, Nakagawara A, Sugimoto T, Inazawa J.: "PPM1D is a potential target for 17q gain in neuroblastoma."Cancer Res. 63. 1876-1883 (2003)
[Publications] Carninci P, Waki K, Shiraki T, Nakagawara A, et al.: "Targeting a complex transcriptome : The construction of the mouse full-length cDNA encyclopedia"Genome Res. 13. 1273-1289 (2003)
[Publications] Nakamura Y, Ozaki T, Koseki H, Nakagawara A, Sakiyama S: "Accumulation of p27^<KIP1> is associated with BMP2-mediated growth arrest and neuronal differentiation of human neuroblastoma-derived cell lines"Biochem.Biophys.Res.Commun.. 307. 206-213 (2003)
[Publications] Ikematsu S, Nakagawara A, Nakamura Y, Sakuma S, Wakai K, et al.: "Correlation of elevated level of serum midkine with poor prognostic factors of human neuroblastomas."Br.J.Cancer. 88. 1522-1526 (2003)
[Publications] Nakagawa T, Takahashi M, Ozaki T, Watanabe K, Hayashi S, Hosoda M, Todo S, Nakagawara A.: "Negative autoregulation of p73 and p53 by ΔNp73 in regulating differentiation and survival of human neuroblastoma cells"Cancer Letters. 197. 105-109 (2003)

2003 Fiscal Year Annual Research Report

小児癌由来cDNAチップを用いた正常個体発生の遺伝子発現ネットワークの網羅的解析

Principal Investigator

大平 美紀 千葉県がんセンター, 生化学研究部, 研究員 (20311384)

Research Products

[Publications] Nakagawara A, Ohira M.: "Comprehensive genomics linking between neural development and cancer : Neuroblastoma as a model. In Special Issue : Neural development and cancer."Cancer Letters. (in press).

[Publications] Ohira M, Morohashi A, Inuzuka H, Nakagawara A, et al.: "Expression profiling and characterization of 4200 genes cloned from primary neuroblastomas : identification of 305 genes differentially expressed between favorable and unfavorable subsets"Oncogene. 22. 5525-5536 (2003)

[Publications] Ohira M, Morohashi A, Nakamura Y, Nakagawara A, et al.: "Neuroblastoma oligo-capping cDNA project : toward the understanding of the genesis and biology of neuroblastoma"Cancer Letters. 197. 63-68 (2003)

[Publications] Kawamoto T, Ohira M, Hamano S, Hori T, Nakagawara A: "High expression of the ovel endothelin-converting enzyme genes, Nbla03145/ECEL1α and β, is associated with favorable prognosis in human neuroblastomas"Int J Oncology. 22. 815-822 (2003)

[Publications] Nakagawara A: "Neural crest development and neuroblastoma : The genetic and biological link"Neuroscience Series, Progress in Brain Research, Elsevier Science Publisher. (in press).

[Publications] Tomizawa M, Watanabe K, Saisho H, Nakagawara A, Tagawa M.: "Down-regulated expression of the CCAAT/enhancer binding protein a and b genes in human hepatocellular carcinoma : a possible prognostic marker"Anticancer Res. 23. 351-354 (2003)

[Publications] Saito-Ohara F, Imoto I, Inoue J, Nakagawara A, Sugimoto T, Inazawa J.: "PPM1D is a potential target for 17q gain in neuroblastoma."Cancer Res. 63. 1876-1883 (2003)

[Publications] Carninci P, Waki K, Shiraki T, Nakagawara A, et al.: "Targeting a complex transcriptome : The construction of the mouse full-length cDNA encyclopedia"Genome Res. 13. 1273-1289 (2003)

[Publications] Nakamura Y, Ozaki T, Koseki H, Nakagawara A, Sakiyama S: "Accumulation of p27^<KIP1> is associated with BMP2-mediated growth arrest and neuronal differentiation of human neuroblastoma-derived cell lines"Biochem.Biophys.Res.Commun.. 307. 206-213 (2003)

[Publications] Ikematsu S, Nakagawara A, Nakamura Y, Sakuma S, Wakai K, et al.: "Correlation of elevated level of serum midkine with poor prognostic factors of human neuroblastomas."Br.J.Cancer. 88. 1522-1526 (2003)

[Publications] Nakagawa T, Takahashi M, Ozaki T, Watanabe K, Hayashi S, Hosoda M, Todo S, Nakagawara A.: "Negative autoregulation of p73 and p53 by ΔNp73 in regulating differentiation and survival of human neuroblastoma cells"Cancer Letters. 197. 105-109 (2003)

大平美紀千葉県がんセンター, 生化学研究部, 研究員 (20311384)