遺伝的揺らぎと孤発癌発生機序解明のための双方向性研究

2009 Fiscal Year Annual Research Report

• Project Area: Molecular Science of Fluctuations toward Biological Functions
• Project/Area Number: 21107509
• Research Institution: Nagoya University
• Principal Investigator: 鈴木 元 Nagoya University, 大学院・医学系研究科, 講師 (80236017)
• Keywords: 揺らぎ / 肺癌 / 細胞周期 / DNA複製 / DNA修復 / チェックポイント / マイクロアレー / ゲノムインスタビリティー

Research Abstract

肺がん検体を用いたmRNAマイクロアレー結果を検討し、孤発癌発生に関与する遺伝的揺らぎ関連遺伝子を抽出した。まず、257種の遺伝子を含むDNA複製、チェックポイント等ゲノムスタビリティーに関わると考えられる揺らぎ関連遺伝子リストを作成した。これを用いてmRNAプロファイル解析を行ったところ、いくつかの遺伝子が肺癌で興味深い発現パターンしていた。このうちPOLD4遺伝子は小細胞癌特異的に発現減少を認め、ノックダウン下で遺伝子の揺らぎであるゲノムインスタビリティー特有の細胞核形態を示した。臨床検体および細胞株中でこの遺伝子変異(SNP)を調べたところ新規のものが一つ発見された。一方、扁平上皮がん特有の発現上昇を認めるものとして、PSMD2を見出した。この遺伝子はこれまで扁平上皮がんでコピーナンバーの増加が認められている領域に存在した。そこで同じ領域に存在しこれまで発癌に深くかかわるとされるPIK3CA遺伝子と発現相関を調べたところ両者には強い相関が観察された。興味深いことに、肺癌の中にはPIK3CAの変化を伴わずにPSMD2のみのコピー数増加を認めるものもあった。また、この遺伝子の発現量と予後の間に有意な相関を認めた。
臨床研究とは反対方向の理論系からの研究として、塩基認識を部分的に欠損するDNA複製遺伝子POLA 1の過剰発現細胞を構築し、DNA複製における塩基挿入ステップの揺らぎ抑制の役割を調べた。このタンパク質は酵素レベルで180倍DNA複製エラーを発生させ、また、細胞レベルでも2-3倍のゲノムインスタビリティーを示した。このことはDNA複製ステップでの揺らぎ抑制機序が細胞の遺伝的特性の維持に重要であることを示している。
以上のように本研究の趣旨である双方向性の研究において、初年度の目標をほぼ達成した。

Research Products

• [Journal Article] Mechanism of Increased PLD1 Gene Expression During Early Adipocyte Differentiation Process of Mouse Cell Line, 3T3-L1 (2010)
  • Author(s): Gao S., et.al
  • Journal Title: J Cell Biochem 109 Pages: 375-382
  • Peer Reviewed
• [Journal Article] ATRA inhibits ceramide kinase transcription in a human neuroblastoma cell line, SH-SY5Y cells : the role of COUP-TFI (2010)
  • Author(s): Murakami M., et.al
  • Journal Title: J Neurochem 112 Pages: 511-520
  • Peer Reviewed
• [Journal Article] Roles of POLD4, smallest subunit of DNA polymerase delta, in nuclear structures and genomic stability of human cells. (2010)
  • Author(s): Huang QM., et.al
  • Journal Title: Biochem Biophys Res Commun 391 Pages: 542-546
  • Peer Reviewed
• [Journal Article] Functions of base selection step in human DNA poly merase alpha. (2010)
  • Author(s): Tanaka S., et.al
  • Journal Title: DNA repair (in press)
  • Peer Reviewed
• [Journal Article] Proteasomal Non-catalytic Subunit PSMD2 as a Potential Therapeutic Target in Association with Various Clinicopathologic Features in Lung Adenocarcinomas. (2010)
  • Author(s): Matsuyama Y., et.al
  • Journal Title: Mol Carcinog (in press)
  • Peer Reviewed
• [Journal Article] p^<27> deregulation by Skp2 overexpression induced by the JAK2V617 mutation (2009)
  • Author(s): Furuhata A., et.al
  • Journal Title: Biochem Biophys Res Commun 383 Pages: 411-416
  • Peer Reviewed
• [Journal Article] GATA-1 and GATA-2 binding to 3' enhancer of WT1 gene is essential for its transcription in acute leukemia and solid tumor cell lines (2009)
  • Author(s): Furuhata A., at.al
  • Journal Title: Leukemia 23 Pages: 1270-1277
  • Peer Reviewed
• [Journal Article] PCNA mono-ubiquitination and activation of translesion DNA polymerases by DNA polymerase alpha (2009)
  • Author(s): Suzuki M., et.al
  • Journal Title: J Biochem 146 Pages: 13-21
  • Peer Reviewed
• [Journal Article] Mutated Ras induced PLD1 gene expression in human colon cancer cell lines and a mouse cell line of NIH3T3 (2009)
  • Author(s): Gao S., et.al
  • Journal Title: Nagoya J Med Sci 71 Pages: 127-136
  • Peer Reviewed
• [Journal Article] Transcriptional regulation of neutral sphingomyelinase 2 gene expression of a human breast cancer cell line, MCF7, induced by anti-cancer drug, daunorubicin. (2009)
  • Author(s): Ito H., et.al
  • Journal Title: Biochim Biophys Acta 1789 Pages: 681-690
  • Peer Reviewed
• [Journal Article] Counterbalance between RB inactivation and miR-17-9 2 overexpression in reactive oxygen species and DNA damage induction in lung cancers (2009)
  • Author(s): Ebi H., et.al
  • Journal Title: Oncogene 28 Pages: 3371-3379
  • Peer Reviewed
• [Journal Article] miRNAの異常と肺癌の分子病態 (2009)
  • Author(s): 細野祥之、鈴木元、高橋隆
  • Journal Title: 実験医学 27 Pages: 1218-1222
• [Presentation] Epigenetic reduction of POLD4/p12 and genomic instability induction in small cell lung cancer." (2009)
  • Author(s): Huang QM., et.al
  • Organizer: The 3^<rd> International Symposium on "Molecular Science of Fluctuations toward Biological Functions
  • Place of Presentation: Nagoya
  • Year and Date: 2009-12-20
• [Presentation] Gene profiling listed a genomic instability gene involved in lung cancer (2009)
  • Author(s): Suzuki M
  • Organizer: Bioregulation of Radiation Response : Damage Recognition, Processing and Cellular Response
  • Place of Presentation: Kyoto
  • Year and Date: 2009-12-01
• [Presentation] Epigenetic reduction of POLD4/p12 and genomic instability induction in small cell lung cancer. (2009)
  • Author(s): Huang QM., et.al
  • Organizer: The 2^<nd> International Symp Nagoya Univ GCOE
  • Place of Presentation: Nagoya
  • Year and Date: 2009-11-25
• [Presentation] Elucidation of Proteasomal Non-catalytic Subunit PSMD2 as Potential Therapeutic Target and Its Co-upregulation with Proteasome Pathway Genes in Association with Various Clinicop athologic Features in Lung Adenocarcinomas. (2009)
  • Author(s): Matsuyama M., et.al
  • Organizer: The 2^<nd> Interntional Symp Nagoya Univ GCOE
  • Place of Presentation: Nagoya
  • Year and Date: 2009-11-25
• [Presentation] Epigenetic reduction of POLD4/p12 and genomic instability induction in small cell lung cancer (2009)
  • Author(s): Huang QM., et.al
  • Organizer: The 19th International Symposium of Hiroshima Cancer Seminar in conjugation with Three Universities' Consortium
  • Place of Presentation: Hiroshima
  • Year and Date: 2009-11-01
• [Presentation] Gene profiling identified a gene involved in genetic instability in lune cancer (2009)
  • Author(s): Suzuki M, Miao HQ, Tomida S, Masuda S, Kamiya K, Osada H, Takashi T.
  • Organizer: 第68回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2009-10-03