| Project/Area Number |
14570140
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Human pathology
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
TOYOKUNI Shinya KYOTO UNIVERSITY, Graduate School of Medicine, Associate Professor, 医学研究科, 助教授 (90252460)
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| Co-Investigator(Kenkyū-buntansha) |
HAGA Hironori KYOTO UNIVERSITY, Graduate School of Medicine, Assistant, 医学研究科, 助手 (10252462)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2002: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | free radicals / oxidative stress / iron / annexin 2 / carcinogenesis / metastasis / peptide nucleic acid / in situ hybridization / 肺癌 / 腎癌 / p15・p16がん抑制遺伝子 / アレル欠損 / in situ hybridization / 鉄トランスポーター / 遺伝子改変動物 |
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| Research Abstract |
Ferric nitrilotriacetate (Fe-NTA) induces oxidative renal damage leading to a high incidence of renal cell carcinoma (RCC) in rats. Differential display analysis of such RCCs revealed elevated expression of annexin 2 (Anx2), a substrate for kinases and a receptor for tissue-type plasminogen adivator and plasminogen. We conducted this study to clarifythe significance of Anx2 in Fenton reaction-based carcinogenesis. Messenger RNA and protein levels of Anx2 were increased time-dependently in the rat kidney after Fe-NTA administration as well as in LLC-PK1 cells after exposure to H_2O_2. The latter was inhibited by pretreatment with N-acetylcysteine, pyrrolidine dithiocarbamate or catalase. immunohistochemistry revealed negligible staining in the normal renal proximal tubules, but strong staining in regenerating proximal tubules, karyomegalic cells and RCCs. Metastasizing RCCs showed higher Anx2 protein levels. Anx2 was phosphorylated at serine and tyrosine residues in these cells and coim
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munoprecipitated with phosphorylated actin. Overexpresison of Anx2 induced a higher cell proliferation rate in LLC-PKI cells. In contrast, a decrease in proliferation leading to apoptosis was observed after Anx2 antisense treatment to cell lines established from Fe-NIA-induced RCCs. These results suggest that Anx2 is regulated by redox status, and that persistent operation of this adaptive mechanism plays a role in the proliferation and metastasis of oxidative stress-induced cancer. Furthermore, we demonstrated that peptide nucleic acid (PNA)-based mRNA in situ hybridization is applicable to paraffin-embedded pathologic specimens produced 50 years ago. We have developed an extremely rapid method for Western blot analysis by the use of low dose of microwave radiation. Finally, we have produced diagnostic criteria for the early humoral rejection in ABO-incompatible liver transplantation. This would be important for investigating the involvement of oxidative stress in the rejection of transplanted liver. Less
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