| Project/Area Number |
09671249
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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 |
General surgery
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| Research Institution | Keio University |
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Principal Investigator |
AIURA Koichi Keio University School of Medicine Instructor, 医学部, 助手 (00184010)
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| Co-Investigator(Kenkyū-buntansha) |
SAITO Junichi Keio University School of Medicine assistant, 医学部, 助手 (30276274)
ITANO Osamu Keio University School of Medicine assistant, 医学部・, 助手 (90265827)
UEDA Masakazu Keio University School of Medicine assistant professor, 医学部, 講師 (50142419)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1998: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1997: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | molecular biology / human genome / two-dimensional electrophoresis / oncogene / RLGS method / ヒト癌 / 癌関連遺伝子 / スクリーニング / クローニング |
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| Research Abstract |
To evaluate alterations of genomic DNAs related to human cancer, we used restriction landmark genomic scanning (RLGS) based on two-dimensional gel electrophoresis. This method has many benefits, one of which can detect thousands of gene fragments on one X-ray film at one time. This analysis was applied for human colorectal cancer and hepatocellular carcinoma. Genomic alterations specific in those cancer tissues were analyzed and the sequences of the gene fragment were identified. Furthermore, we have clarified the correlation between those genomic alterations and clinical characteristics.
1 . Average twelve changes in DNA spots after two-dimensional gel electrophoresis were seen in a case with colorectal cancer. one of the DNA fragments in the spot was extracted and the nucleotide sequence was determined by ABI PRISM GENETIC ANALYSER.The sequence (410 bps) was found to be highly homologous to human fatty acid syhthase (65 %) in Gene Bank database. The expression of the RNA was also observed in the only cancer tissues by northern blot hybridization using the probe from the DNA spot.
2. Average sixteen changes in DNA spots were seen in a case with hepatocellular carcinoma. The survival rates for patients with <greater than or equal> 16 changed RLGS spots were significantly lower than those for patients with fewer changed RLGS spots ( <less than or equal> 15 spots). Four spots of the DNA alterations of cancer tissues were extracted and sequenced. One (1088 bps) was located at chromosome 8q21 and corresponded to human tandem repeat sequence. Another one (1415 bps) was centromeric NotI cluster. The disease-free survival rates for patients with hepatocellular carcinoma were closely associated with the intensity of those
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