Herbergs J, Arends J W, Bongers E M, Ramaekers F C, Hopman A H
Department of Pathology, University Hospital Maastrict, The Netherlands.
Genes Chromosomes Cancer. 1996 Jun;16(2):106-12. doi: 10.1002/(SICI)1098-2264(199606)16:2<106::AID-GCC4>3.0.CO;2-3.
In this study, we demonstrated the clonal origin of trisomy for chromosome 7 in epithelial cells of colon neoplasia. By using the double-target fluorescence in situ hybridization (FISH) technique in frozen tissue sections that were also immunostained for keratin and vimentin, ratio analysis of FISH signals for chromosomes 7 and 17 could be performed in epithelial (cytokeratin-positive) or stromal (vimentin-positive) areas. The data demonstrated that trisomy for chromosome 7 is found exclusively in the epithelial compartments and not in the stroma of colon adenocarcinoma. We then demonstrated the occurrence of trisomy for chromosome 7 in the different types of epithelial neoplastic cells, i.e., columnar and goblet cells, which were isolated from frozen tissue sections by mechanical disaggregation of colon tissue and mild lysis of the cells while protease activity was inhibited. In these cell suspensions, the columnar cells were detected with an antibody to villin, and the goblet cells were stained for mucin, whereas all cells were subsequently subjected to FISH for chromosome 7. For analysis of neuroendocrine cells, which are present in a very low frequency in colon neoplasia, frozen tissue sections that were immunostained for Chromogranin A could be used. Individual neuroendocrine cells could be distinguished in these thin frozen tissue sections. The presence of trisomy for chromosome 7 in all three different epithelial cell types strengthens our suggestion that this chromosomal aberration is found in the epithelial stem cell compartment of colon neoplasia.
在本研究中，我们证实了结直肠癌上皮细胞中7号染色体三体的克隆起源。通过在冷冻组织切片中使用双靶点荧光原位杂交（FISH）技术，同时对细胞角蛋白和波形蛋白进行免疫染色，可在上皮（细胞角蛋白阳性）或基质（波形蛋白阳性）区域进行7号和17号染色体FISH信号的比值分析。数据表明，7号染色体三体仅在上皮区室中发现，而在结肠腺癌的基质中未发现。然后，我们证实在不同类型的上皮肿瘤细胞，即柱状细胞和杯状细胞中存在7号染色体三体，这些细胞是通过对结肠组织进行机械解离并在抑制蛋白酶活性的同时对细胞进行轻度裂解，从冷冻组织切片中分离出来的。在这些细胞悬液中，用抗绒毛蛋白抗体检测柱状细胞，用黏蛋白对杯状细胞进行染色，随后对所有细胞进行7号染色体的FISH检测。对于结肠肿瘤中频率极低的神经内分泌细胞的分析，可使用对嗜铬粒蛋白A进行免疫染色的冷冻组织切片。在这些薄的冷冻组织切片中可以区分单个神经内分泌细胞。在所有三种不同上皮细胞类型中均存在7号染色体三体，这进一步支持了我们的观点，即这种染色体畸变存在于结肠肿瘤的上皮干细胞区室中。
