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多色荧光原位杂交技术是在常规荧光原位杂交技术基础上建立的一种染色体分析手段,已广泛应用于肿瘤遗传学的研究,尤其是用于染色体隐匿性重排及复杂核型分析。本研究旨在利用现有实验条件,建立一种可靠的人类全染色体分析方法———重复多色荧光原位杂交技术(repetitive multicolour fluorescence in situ hybridization,RM-FISH)。采用特异的全染色体涂染探针,分别用Cy3、Cy5、FITC3种荧光素直接标记,组合制备4组六色的探针池,对同一染色体片先后进行4次杂交,通过装备了4组滤镜的荧光显微镜捕获信号。RM-FISH技术结合反相DAPI显带对1例原发性食管鳞癌进行核型分析,获得了全部染色体的特异信号。本研究建立的RM-FISH简便、易行,是光谱核型分析的一种较理想的替代技术。
Multi-color fluorescence in situ hybridization is a chromosomal analysis method established on the basis of routine fluorescence in situ hybridization. It has been widely used in tumor genetics research, especially for the occult chromosomal rearrangement and complex karyotyping. The purpose of this study is to establish a reliable method for human chromosome analysis by using the existing experimental conditions --- repetitive multicolor fluorescence in situ hybridization (RM-FISH). Using a specific chromosome-coated probe, respectively, with Cy3, Cy5, FITC three kinds of fluorescein direct labeling, the combination of preparation of six six-color probe pool, the same chromosome four times in succession, equipped with four filters A fluorescence microscope captures the signal. RM-FISH combined with reverse-phase DAPI banding karyotype analysis of a case of primary esophageal squamous cell carcinoma, obtained all the chromosome-specific signals. The RM-FISH established in this study is simple and easy to perform and is an ideal substitute for spectral karyotyping.