細胞周期関連分子Cdk6による骨代謝調節機構の解明-遺伝子操作マウスの解析-

• Project/Area Number: 17791438
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Surgical dentistry
• Research Institution: The University of Tokyo
• Principal Investigator: 小笠原 徹 The University of Tokyo, 医学部附属病院, 講師 (20359623)
• Project Period (FY): 2005 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2007: ¥1,000,000 (Direct Cost: ¥1,000,000); Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000); Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000)
• Keywords: 分化 / 細胞周期 / 骨代謝 / Cdk6 / 骨軟骨

Research Abstract

本年度は、細胞周期関連分子Cdk6による骨代謝調節機構の解明を目的にCdk6ノックアウトマウスの骨組織を解析した。まず生後8週から12週齢のCdk6ノックアウトマウスと同胞野生型マウスの全身での表現系ならびに骨組織を比較した。その結果、Cdk6ノックアウトマウスでは時期特異的に分化能の変化が生じることが明らかとなった。そこで、そのメカニズムを詳細に分析するために骨細胞の分化能をRealtime-RT-PCRと組織形態学的手法によって解析するとともに、その増殖能をXTTアッセイで評価した。また、骨芽細胞様細胞MC3T3-E1を用いた研究成果によって、Cdk6ならびにCdk4が骨形成に必須の転写因子Runx2と結合し、Cdk4-Runx2複合体がOC promoterに結合する一方、Cdk6-Runx2複合体はOC promoterには結合しないことが明らかとなった。すなわち、(1)Cdk6がRunx2と結合してpromoterへのRunx2の結合を選択的に阻害することでその転写活性を抑制すること、(2)Cdk4がRunx2と結合して転写共役因子として骨芽細胞分化を促進することが示された。この結果に基づいてCdk6ノックアウトマウス由来骨芽細胞内におけるRunx2と各種分化増殖制御因子との相関を検討したところ、Cdk6ノックアウトマウス由来骨芽細胞では、Runx2の発現やその細胞内での局在自体は野生型マウス由来骨芽細胞と比べて差はないが、いくつかの分化制御因子の発現パターンや機能に差があることが示唆された。(656字)

Research Products

• [Journal Article] Effects of transforming growth factor (TGF)-β1 in combination with fibroblast growth factor (FGF)-2 and insulin-like growth factor (IGF)-I on chondrocytes proliferation culture for the cartilage regenerative medicine. (2009)
  • Author(s): Ogasawara T et al.
  • Journal Title: Asian J Oral Maxillofac Surg(in press)
  • Peer Reviewed
• [Journal Article] Aptitude of Auricular and Nasoseptal Chondrocytes Cultured under a Monolayer or Three-Dimensional Condition for Cartilage Tissue Engineering. (2009)
  • Author(s): Asawa Y et al.
  • Journal Title: Tissue Eng Part A.(in press)
  • Peer Reviewed
• [Journal Article] C/EBPβ promotes transition from proliferation to hypertrophic differentiation of chondrocytes through transactivation of p57Kip2. (2009)
  • Author(s): Hirata M et al.
  • Journal Title: PLoS ONE(in press)
  • Peer Reviewed
• [Journal Article] Selection of highly osteogenic and chondrogenic cells from bone marrow stromal cells in biocompatible polymer-coated plates. (2009)
  • Author(s): Liu G et al.
  • Journal Title: J Biomed Master Res A(in press)
  • Peer Reviewed
• [Journal Article] Phosphorylation of GSK-3beta by cGMP-dependent protein kinase II promotes hypertrophic differentiation of murine chondrocytes. (2008)
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  • Journal Title: J Clin Invest 118 Pages: 2506-2515
  • Peer Reviewed
• [Journal Article] Patchedl haploinsufficiency increases adult bone mass and modulates Gli3 repressor activity. (2008)
  • Author(s): Ohba S et al.
  • Journal Title: Dev Cell 14 Pages: 689-699
  • Peer Reviewed
• [Journal Article] Growth factor contents of autologous human sera prepared by different production methods and their biological effects on chondrocytes. (2008)
  • Author(s): Tanaka Y et al.
  • Journal Title: Cell Biol Int 32 Pages: 505-514
  • Peer Reviewed
• [Journal Article] Three-Dimensional Microenvironments Retain Chondrocyte Phenotypes During Proliferation Culture (2007)
  • Author(s): Tsuguharu Takahashi et al.
  • Journal Title: Tissue Engineering (in press)
• [Journal Article] Cartilage tissue engineering using human auricular chondrocytes embedded in different hydrogel materials. (2006)
  • Author(s): Hisayo Yamaoka et al.
  • Journal Title: J Biomed Mater Res A 78 Pages: 1-11
• [Journal Article] Cartilage tissue engineering using human auricular chondrocytes embedded in different hydrogel materials. (2006)
  • Author(s): Hisayo Yamaoka et al.
  • Journal Title: J Biomed Mater Res (in press)
• [Journal Article] Inhibition of Cdk6 expression through p38 MAP kinase is involved in differentiation of mouse prechondrocyte ATDC5. (2005)
  • Author(s): Toru Moro et al.
  • Journal Title: J Cell Physiol 204 Pages: 927-933
• [Journal Article] Mutation in cGMP-dependent protein kinase II causes dwarfism in a rat mutant KMI through uncoupling of proliferation and differentiation of chondrocytes. (2005)
  • Author(s): Hirotaka Chikuda et al.
  • Journal Title: J Bone Miner Metab 23 Pages: 200-204
• [Journal Article] Synergistic effects of FGF-2 with insulin or IGF-I on the proliferation of human auricular chondrocytes. (2005)
  • Author(s): Tsuguharu Takahashi et al.
  • Journal Title: Cell Transplant 14 Pages: 683-693
• [Journal Article] Suppression of Adjuvant-Induced Arthritic Bone Destruction by Cyclooxygenase-2 Selective Agents with and without Inhibitory Potency against Carbonic Anhydrase II. (2005)
  • Author(s): Mika Katagiri et al.
  • Journal Title: J Bone Miner Res 21 Pages: 219-227
• [Presentation] 間葉系細胞の分化多能性維持機構におけるNanogの役割 (2008)
  • Author(s): 小笠原徹, ほか
  • Organizer: 第26回日本骨代謝学会学術集会
  • Place of Presentation: 大阪府大阪市
• [Presentation] C/EBPβ/p57Kip2 signaling maintains transition from proliferation to hypertrophic differentiation of chondrocytes during skeletal growth. (2008)
  • Author(s): Hirata M et al.
  • Organizer: 30th Annual Meeting of the American Society for Bone and Mineral Research.
  • Place of Presentation: Montreal, Quebec, Canada
• [Presentation] C/EBP-beta promotes hypertrophic differentiation of chondrocytes through transactivation of p57Kip2 during endochondral ossification. (2008)
  • Author(s): Kugimiya F et al.
  • Organizer: 2008 World Congress on Osteoarthritis
  • Place of Presentation: Rome, Italy
• [Remarks]
  • URL: http://plaza.umin.ac.jp/~oralsurg/