2001 Fiscal Year Final Research Report Summary
Analysis of stress defense mechanism against oral cancer by using A170 gene knockout mouse
| Project/Area Number |
11470429
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Surgical dentistry
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| Research Institution | University of Tsukuba |
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Principal Investigator |
YOSHIDA Hiroshi Univ. of Tsukuba, Inst. Of Clinical medicine, professor, 臨床医学系, 教授 (80014330)
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| Co-Investigator(Kenkyū-buntansha) |
YUSA Hiroshi Univ. of Tsukuba, Inst.of Clinical medicine, instructor, 臨床医学系, 助手 (40292560)
ONIZAWA Kojiro Univ. of Tsukuba, Inst.of Clinical medicine, assistant professor, 臨床医学系, 講師 (60194578)
ISHII Tetsuro Univ. of Tsukuba, Inst.of Basic medicine, associate professor, 基礎医学系, 助教授 (20111370)
YANAGAWA Toru Univ. of Tsukuba, Inst.of Clinical medicine, assistant professor, 臨床医学系, 講師 (10312852)
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| Project Period (FY) |
1999 – 2001
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| Keywords | A170 / zeta interacting protein / gene targeting / oxidative stress / stress response |
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| Research Abstract |
In this study we focused on the oxidative stress protein A170 which was estimated to play a important role in oral and maxillofacial lesions because it was associated with the oncogene and osteoblast differentiation. We planed 1) to construct A170 gene knockout mouse and 2) to analyze the stress : response of the oxidative stress proteins including A170 from clinical and basic viewpoints.
1) About knockout mouse construction we carried out the following procedures. We isolated the genomic clone and characterized the gene of A170 from BAC/129Sv mouse genome library. After mapping we designed the A170-null targeting vector. The modified gene with the vector was introduced into the ES cells by electroporation. Genetically altered cells were injected into embryonic blastocysts derived from C57/B6N and then implanted into a surrogate mother. Chimeras were generated with a high agouti percentage coat which indicated acceptance of the gene mutation. These chimeric mice were then mated with CS7/B6 mice. The F1 offspring containing the agouti coat indicated germline transmission. The germline transmission was confirmed by FCR. Finally we mated Flmouse and knock out mouse (F2) construction was completed.
2) While knockout mouse construction we investigated the induction of A170 by stress agents using the rat. We also investigated Peroxiredoxin I (Prx I = MSP23) and the other stress inducible protein expression as a foundation for A170 knockout-mouse analysis. Moreover, we also investigated the expression level of oral cancer from clinical samples.
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