2006 Fiscal Year Final Research Report Summary
Development of novel screening method by detection of APC mutation from colorectal cancer cells in stool
| Project/Area Number |
17591404
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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 |
Digestive surgery
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
IKEDA Satoshi Hiroshima University, Graduate School of Biomedical Science, Faculty member, 大学院医歯薬学総合研究科, 寄附講座教員 (60397924)
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| Co-Investigator(Kenkyū-buntansha) |
OKAJIMA Masazumi Hiroshima University, Graduate School of Biomedical Science, Faculty member, 大学院医歯薬学総合研究科, 寄附講座教員 (90274068)
ASAHARA Toshimasa Hiroshima University, Hospital, Professor, 病院・教授 (70175850)
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| Project Period (FY) |
2005 – 2006
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| Keywords | Stool test / APC / MCR / Stop codon / Yeast / URA3 / 5FOA |
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| Research Abstract |
We noticed that 80-90% of colorectal cancer have APC (adenomatous polyposis coli) gene mutations. We aimed to detect the presence of cancer extracting DNA of fallen cancer cells in stool and using yeast in this project.
1. Cell line from human colorectal cancer : SW480 has mutation that causes stop codon in MCR (mutation cluster lesion) of APC gene. We amplified MCR lesion of APC gene of normal DNA from lymphocyte and mutated DNA from SW480 and caused yeast transformation after transducing into URA3(+) vector. Then we tested colony formation of yeast in normal and 5FOA(+) culture medium. Colony formation was detected in yeast including APC from SW480 due to stop codon of APC mutation coursing absence of URA3, while colony was not seen in yeast including APC from normal DNA
2. Next, we added further improvement concerning its amount and method on established technique of DNA extraction from stool. Stable DNA extraction was successfully obtained.
3. We have established the technique of DNA extraction from stool of normal volunteer, per of APC MCR, transformation of yeast. However, further improvement of efficiency of DNA transformation is still needed. For cancer patient, technique of DNA sequence of APC from samples has been completed. Now we are planning a gene analysis consulting hospital ethics committee
4. Now we have not applied a patent because this study is not mature enough. We are planning the application with further examination.
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