The trial for chromosomal distribution analysis of in situ PCR and FISH signals as an index of malignamcy in gastro-enterological carcinomas

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Concerning inherent karyotypic heterogeneities of solid tumor, we evaluated both the exhibiting number of chromosome 17 and nuclear DNA content on an identical nucleus by multi-parametric analysis in order to demonstrate the alteration in number of chromosome 17 among cytofluorometrically distinct subpopulations.In the present study concerning colorectal carcinoma, we have collected both of these values sequentially on an identical nucleus by using computer controlled auto-scanning stage.1) We investigated 8 lesions of surgically resected colorectal carcinomas, which were classified as aneuploid in quantitative DNA analysis and also exhibited an increase of 17-aneusomy nuclei. These observations indicate taht the distribution of number of chromosome 17 reflects an endoreduplication of genome content, yet, it does not alter in accordance with the phase of cell cycle.2) We introduced the thermal cycler for in situ PCR in 1996. the conditions of in situ PCR method were evaluated such as sample preparations primer selection, condition setting and labeling mmethod of signal.3) It is necessary to evaluate nuclear DNA content simultaneously in order to assess an essential cytogenetic charge. Althoug both nuclear DNA content and number of a particular chromosome should be evaluated simultaneously on an identical nucleus, we usually could not obtain a successful result, because the denaturation process, which is required in FISH procedure, hinders correct evaluation of nuclear DNA content.4) We still have the problem concerning in situ PCR method applying solid tumor such as colorectal carcinomas.