| Project/Area Number |
06670512
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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 |
Gastroenterology
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| Research Institution | Hirosaki University |
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Principal Investigator |
SAITO Hiroshi Hirosaki University, School of Medicine, lecturer, 医学部, 講師 (70196004)
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| Co-Investigator(Kenkyū-buntansha) |
OOKAWA Keizou Hirosaki University, School of Medicine, assistant, 医学部, 助手 (70250206)
吉田 豊 弘前大学, 医学部, 教授 (80003375)
中嶋 均 弘前大学, 医学部, 助手 (20227797)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1996: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1995: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1994: ¥700,000 (Direct Cost: ¥700,000)
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| Keywords | colorectal cancer / metastasis / cell line / genetic alteration / CAR / KAI1 / Smad 4 / DNA polymerase beta / KAII / RT-PCR / 大腸がん |
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| Research Abstract |
For the screening of candidate gene regulating metastatic potential of colorectal cancer, we established the system, by which genetic alterations in small samples such as biopsy specimen can be analyzed easily and rapidly by using fluorescein-labeled PCR method, SSCP analysis and multiplex RT-PCR.Alterations of the DCC and p53 genes were detected in 10 of 11 colorectal cancer. Therefore, the samples were properly obtained from cancer tissues and the system was available. In contrast, mutation of DNA polymerase beta mRNA was not detected in our samples including primary tumor, liver metastasis and cell lines. The result was different from the previous report that its mutation was detected in 80% of advanced colorectal cancer. We also investigated correlation of metastatic potential with expression of the KAI1 and CAR genes, known as a candidate metastatic suppressor genc, and the Smad 4 gene, located on chromosome 18q21 near the DCC gene. Their expression was quantitated using beta-actin expression as internal control in 12 colorectal adenomas, 38 primary carcinomas, 10 liver metastases and 7 cell lines. The results showed significant correlation of tumor progression with down-regulation of Smad4 and up-regulation of CAR.On the other hand, KAI1 expression was not significantly associated with tumor progression but liver metastases were divided into two groups according to KAI1 expression ; increased expression and decreased expression.
We have been trying to establish a pair of cell lines from primary tumor and liver metastasis from the same patient. At present, two cell lines were obtained from liver metastases but further efforts are needed. Isolation of candidate metastasis-associated genes is also continued by the arbitrary-primed PCR method and the gene display method with fluorescein labeling. We will perform further studies for identification of the gene-regulating metastatic potential of colorectal carcinoma based on the above progress in this project.
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