Shroyer K R, Gudlaugsson E G
Department of Pathology, University of Colorado, Health Sciences Center, Denver 80262-0216.
Hum Pathol. 1994 Mar;25(3):287-92. doi: 10.1016/0046-8177(94)90201-1.
Clonality of archival formalin-fixed tissue sections was analyzed by polymerase chain reaction amplification of a portion of the X-linked phosphoglycerate kinase (PGK-1) gene. Amplification was successful in 29 of 36 cases of uterine endometrioid adenocarcinoma. Five of these cases, including both tumor and control tissue from the same patients, were heterozygous for the BstXI polymorphic site of the PGK-1-amplified product, permitting analysis of clonality. Pretreatment of the DNA with HpaII blocked amplification of one of the two PGK-1 alleles from four of five cases of tumor, indicating the clonal pattern of X chromosome inactivation in these cases. In contrast, in DNA from paired control tissues HpaII pretreatment had no effect, indicating a random pattern of X chromosome inactivation in normal tissue. One of the cases of endometrioid adenocarcinoma contained a high proportion (45%) of nontumor cells, precluding the determination of clonality. We conclude that polymerase chain reaction amplification can be used for the determination of the pattern of X chromosome inactivation in formalin-fixed tissue sections. Such an approach makes it feasible to include specimens from archival tissue collections in the analysis of clonality.
通过对X连锁磷酸甘油酸激酶（PGK-1）基因的一部分进行聚合酶链反应扩增，分析存档福尔马林固定组织切片的克隆性。在36例子宫内膜样腺癌病例中，有29例扩增成功。其中5例，包括来自同一患者的肿瘤组织和对照组织，PGK-1扩增产物的BstXI多态性位点为杂合子，从而可以进行克隆性分析。用HpaII对DNA进行预处理可阻断5例肿瘤病例中两PGK-1等位基因之一的扩增，表明这些病例中X染色体失活的克隆模式。相反，在配对对照组织的DNA中，HpaII预处理没有效果，表明正常组织中X染色体失活是随机模式。其中1例子宫内膜样腺癌含有高比例（45%）的非肿瘤细胞，无法确定克隆性。我们得出结论，聚合酶链反应扩增可用于确定福尔马林固定组织切片中X染色体失活的模式。这种方法使得将存档组织库中的标本纳入克隆性分析成为可能。
