广泛标记系统在石蜡包埋膀胱癌组织比较基因组杂交中的应用

目的 探讨广泛标记系统（ULS）在石蜡包埋组织比较基因组杂交（CGH）中的应用价值,研究膀胱癌石蜡包埋组织中染色体基因组非平衡性情况.方法 对比实验：正常膀胱黏膜组织基因组DNA,用DNA酶进行酶切至适当大小的片段作为探针,用广泛标记系统直接标记DNA探针,并按1：1的量与经缺口平移标记的同一个组织的DNA探针进行CGH.提取膀胱移行细胞癌石蜡包埋组织中基因组DNA,经测定浓度、纯度,并经琼脂糖电泳筛选出20例适合于CGH的标本.同法进行ULS标记,并与缺口平移法标记的正常参照DNA进行CGH.结果 对比实验中,同一个正常膀胱黏膜组织应用2种标记方法进行CGH后,荧光强度相同,不需要调整探针的量.在所有膀胱癌组织中均检测到基因组DNA的畸变,其中发生频率较高的染色体增益区段或位点位于1q、3q、5p、8q、17q;缺失区段或位点位于8p、9q、11q、11p、17p.结论 膀胱癌中存在染色体基因组的不平衡.应用石蜡包埋的组织进行间接法CGH时,ULS标记系统是一种非常有价值的标记方法.这为充分利用石蜡档案进行CGH的研究提供了良好的方法.

The Application of Universal Linkage System in Comparative Genomic Hybridization of Formalin-fixed, Paraffin-embedded Bladder Cancer Tissues

Purpose To investigate the application value of universal linkage system (ULS) in compa- rative genomic hybridization (CGH) of formalin-fixed, paraffin-embedded tissue. Methods In contrast test, normal bladder mucosal tissues were selected to extract genomic DNA, and used the DNase plus ULS to label the tumor probes. Same amount of probes were used to perform CGH with that of labeled by nick translation labeling method in order to adjust the quantity of the probes.A total of 20 cases of formalin-fixed, paraffin-embedded bladder cancer specimens were selected by their concentration, purity and fragments.And then, every specimen was detected by CGH technique using the same method, and the aberrations of chromosome segments or loci were got by statistics analysis. Results In the contrast test, the density of the chromosomal fluorescence was the same, the CGH protocol could be performed without adjus- ting the amount of probes. Among all the bladder cancer tissues tested, there existed chromosomal aberrations, the most frequent chromosomal gains were 1p,3p,5p,8q,17q. However, the most frequent chromosomal loss were 8p,9q,11q,11p,17p. Conclusions There were chromosomal genomic imbalances in the bladder cancer tissues tested. The ULS labeling system could be used in formalin-fixed, paraffin-embedded tissues to perform CGH. This will give us a new way in full use of archival tissues in CGH.