van Dekken H, Kerstens H M, Tersteeg T A, Verhofstad A A, Vooijs G P
Department of Pathology, Erasmus University Rotterdam, The Netherlands.
J Pathol. 1992 Nov;168(3):317-24. doi: 10.1002/path.1711680312.
In this study, non-isotopic in situ hybridization (ISH) was used for the cytogenetic and histological examination of urological (prostatic adenocarcinoma) and endocrine (phaeochromocytoma) tumour cell nuclei in 4 microns paraffin-embedded tissue sections. In order to investigate preservation of tissue morphology, standard heat denaturation was compared with a mild enzymatic treatment for the production of single-stranded (ss)-DNA for ISH. Numerical analysis by ISH with chromosome-specific repetitive DNA probes for chromosomes 1, 7, and 11 revealed overrepresentation of chromosome 7 in the phaeochromocytoma (P < 0.01). The constitutional underrepresentation of the Y chromosome was easily detected in the prostate tumour (P << 0.01) when probed for chromosomes 7, 16, and Y. The enzymatic treatment appeared superior to heat denaturation with respect to tissue architecture in the phaeochromocytoma, while no clear difference was observed in the prostatic cancer. ISH probe patterns were similar for the two types of denaturation in both tumours (P > or = 0.20). We conclude that (1) ISH can be used for the identification of numerical cytogenetic changes in solid tumour cell nuclei within archival tissue sections; and (2) mild 'denaturation' protocols, replacing heat, are preference in retaining tissue architecture in fragile tumour specimens.
在本研究中,采用非同位素原位杂交(ISH)技术,对4微米石蜡包埋组织切片中的泌尿生殖系统(前列腺腺癌)和内分泌系统(嗜铬细胞瘤)肿瘤细胞核进行细胞遗传学和组织学检查。为了研究组织形态的保存情况,将标准热变性与温和酶处理进行比较,以制备用于ISH的单链(ss)DNA。使用针对1号、7号和11号染色体的染色体特异性重复DNA探针进行ISH数值分析,结果显示嗜铬细胞瘤中7号染色体过度表达(P < 0.01)。当用7号、16号和Y染色体探针检测时,在前列腺肿瘤中很容易检测到Y染色体的体质性低表达(P << 0.01)。就嗜铬细胞瘤的组织结构而言,酶处理似乎优于热变性,而在前列腺癌中未观察到明显差异。两种肿瘤中两种变性类型的ISH探针模式相似(P ≥ 0.20)。我们得出结论:(1)ISH可用于识别存档组织切片中实体肿瘤细胞核的数值细胞遗传学变化;(2)在保留脆弱肿瘤标本的组织结构方面,取代热变性的温和“变性”方案更受青睐。