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Analysis of molecular mechanism and global gene expression on urolithiasis using cDNA array

Research Project

Project/Area Number 13671678
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Urology
Research InstitutionKANAZAWA MEDICAL UNIVERSITY

Principal Investigator

MIYAZAWA Katsuhito  KANAZAWA MEDICAL UNIVERSITY, UROLOGY, Instructor, 医学部, 講師 (60219772)

Co-Investigator(Kenkyū-buntansha) SUZUKI Koji  KANAZAWA MEDICAL UNIVERSITY, UROLOGY, Professor, 医学部, 教授 (70064615)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2001: ¥2,300,000 (Direct Cost: ¥2,300,000)
Keywordsurolithiasis / Calcium oxalate / CDNA array / Gene expression / Renal epithelial cell / Potassium oxalate / DNAマクロアレイ
Research Abstract

Purpose :
Kidney stone formation is a complex process, and numerous genes participate in this cascade. The binding and internalization of calcium oxalate monohydrate (CON) crystals, the most common crystal in renal stones by renal epithelial cells may be a critical step leading to kidney stone formation. Exposure to CON crystals alters the expression of various genes, but previous studies on gene expression have generally been limited. To obtain more detailed insight into gene expression, we examined gene expression profiles in renal epithelial cells exposed to CON crystals using cDNA macroarray. Materials and Methods : NRK-52E cells were exposed to CON crystals for 60 and 120 minutes. Poly (A)^+ RNA was isolated and converted into ^<32>P-labeled first-strand cDNA, then the cDNA probe was hybridized to the membrane. Hybridization images were scanned and the signal intensities were quantified. Expression of mRNA of 1176 genes were analyzed with global sum normalization methods.
Results :
E … More xposure to CON crystals altered the expression of some of the genes reported previously. Furthermore, novel genes were also identified. The genes for 35 proteins were altered in the expression level by more than 2-fold : annexin V, p53-binding mouse double minute 2 homolog, osteopontin, fibronectin, steroidogenic acute regulatory protein, clusterin, coagulation factor II (thrombin) receptor, mitogen-activated protein kinase 3, cathepsin B, cathepsin L, plasminogen activator inhibitor 1, cytokeratin 8, vimentin, hydrophobic surfactant associated protein C, inhibitor of DNA binding 1, 7-dehydrocholesterol reductase, cytochrome P-45O 2C23, myeloid cell differentiation protein 1, high mobility group protein 2, 5-hydroxytryptamine receptor 5B receptor, cytochrome c expressed in somatic tissues, connective tissue growth factor, tissue inhibitor of metalloproteinase 1, V(D)J recombination activating protein 1, Tclone15, thymosin beta 10, junD proto-oncogene, inhibitor of DNA binding 2, inhibitor of DNA-binding protein 3, G1/S-specific cyclin D1, collagen, insulin-like growth factor binding protein 2, insulin-like growth factor-binding protein 6, profilin 1 and metallothionein 1. Twenty genes had altered expression after a 60-minute exposure to COM crystals. Thirteen genes were up-regulated and seven genes were down-regulated. Nineteen genes had altered expression after a 120-minute exposure to COM crystals. Nine genes were up-regulated and ten genes were down-regulated.
Conclusions :
We performed a large-scale analysis of gene expression in renal epithelial cells exposed to COM crystals, and identified the genes differentially regulated. cDNA macroarray is a useful tool for evaluating gene expression in urolithiasis research. Less

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] 宮澤 克人: "CaOx結晶による腎尿細管細胞の遺伝子発現について-DNAマクロアレイによる解析"日本尿路結石症学会誌. 1. 114-116 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 宮澤 克人: "尿路結石再発予防の薬物療法"泌尿器外科. 16. 1157-1163 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 宮澤 克人: "尿路結石に対するアルコールの影響について"日本尿路結石症学会誌. 2. 92-95 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 宮澤 克人: "尿路結石と飲水および飲料物について"泌尿器科紀要. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 相原 衣江: "ヒト腎尿細管培養細胞を用いた尿路結石関連蛋白質の発現についての検討"日本尿路結石学会 第11回学術集会記録集. 11. 183-186 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 森山 学: "蓚酸による腎尿細管細胞障害における活性酸素発生部位に関する検討"日本尿路結石学会 第11回学術集会記録集. 11. 187-190 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] KATSUHITO MIYAZAWA: "Gene expression on renal epithelial cells by CaOx crystal. -analysis with cDNA array-"Japanese Society on Urolithiasis Research. 1. 114-116 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] KATSUHITO MIYAZAWA: "The preventive pharmacological treatment of patients with calcium stone disease"Japanese Journal of Urological Surgery. 16. 1157-1163 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] KATSUHITO MIYAZAWA: "The influence of alcohol consumption on the risk of kidney stones"Japanese Society on Urolithiasis Research. 2. 92-95 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] KATSUHITO MIYAZAWA: "Study of beverage and kidney stone disease"Acta Urologica Japonica. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] KINUE AIHARA: "Expression of urolithiasis related protein on human renal epithelial cells"Japanese Society on Urolithiasis Research. 11. 183-186 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] MANABU T MORTYAMA: "Localization of superoxide in oxalate-assosiated injury to HK-2 cells"Japanese Society on Urolithiasis Research. 11. 187-190 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 宮澤 克人: "CaOx結晶による腎尿細管細胞の遺伝子発現について-DNAマクロアレイによる解析"日本尿路結石症学会誌. 1・1. 114-116 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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