Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1996: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1995: ¥1,600,000 (Direct Cost: ¥1,600,000)
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Research Abstract |
A number of genetic alterations have been detected to occur in human malignancy during multistep progression. Metastatic lesion, therefore, seems to be accompanied by genetic changes in addition to those occurred in the primary lesion. The critical genes related to the tumor metastasis should be identified to elucidate the genetic mechanism for the metastasis and to distinguish between the double primaries and the metastatic tumors. We investigated the genetic event closely related to the tumor metastasis or invasion using arbitrarily primed polymerase chain reaction (AP-PCR) as a DNA fingerprinting analysis in the tumor with multiple sites. In malignancy of female genital tract, the tumor often have multiple lesions, which are not known double primary ones or metastatic tumor from the primary. We extracted DNA of the multiple tumors and the corresponding normal tissue from 8 patients with simultaneous occurring ovarian and endometrial cancers, 8 patients with disseminated ovarian canc
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er and 5 patients with endometrial cancers with metastatic lesions. Our PCR had a lower stringency of annealing temperature at 37゚C to 42゚C in the first 5 cycles of the PCR reaction. One AP-PCR trial made 15 to 25 electrophoretic bands with each PCR primer. Twenty PCR primers and their sequence were randomly chosen. We failed to examine the clonality using AP-PCR-single strand conformation polymorphism analysis because of unclear interpretation of the result. Tumor DNA of every patients showed at least one abnormal band in comparison of normal DNA in the AP-PCR analysis. Electrophoretic band pattern was identical among multiple tumor DNA in 8 patients with disseminated ovarian cancers, 5 patients with metastatic endometrial cancer and 7 patients with simultaneous occurring ovarian and endometrial cancer. These tumors were considered to be identical origin, and therefore, these were primary and metastasis. On the other hand, one of 8 patients with ovarian and endometrial cancer showed different pattern of DNA band in three lesions, and being considered as primary and metastasis. These results of tumor clonality were same as those from clinicopathological analysis. The AP-PCR is useful for analysis of clonality of the gynecological tumor, although metastasis specific DNA change was not identified in this study. Less
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