所谓肺硬化性血管瘤组织中两种主要细胞的雄激素受体、磷酸甘油酸酯激酶基因多态性分析

Study of androgen receptor and phosphoglycerate kinase gene polymorphism in major cellular components of the so-called pulmonary sclerosing hemangioma

目的探讨所谓肺硬化性血管瘤(PSH)组织内的多角形细胞和表面立方细胞的克隆性组成及意义。方法选PSH患者手术切除标本19例,其中女性17例。石蜡切片HE染色,应用激光捕获显微切割技术获取PSH组织中多角形细胞和表面立方细胞,分别提取基因组DNA,用甲基化敏感的限制性内切酶HhaⅠ或HpaⅡ消化,巢式聚合酶链反应扩增X染色体连锁的雄激素受体(AR)基因及磷酸甘油酸酯激酶(PGK)基因。AR基因产物经变性聚丙烯酰胺凝胶电泳、银染显示其长度多态性;PGK基因产物经BstⅪ消化后,琼脂糖电泳显示其酶切位点的多态性。结果17例女性PSH组织,扩增成功AR基因及PGK基因的多态性分别为53%(8/15)和27%(4/15)。对适合于检测的10例标本分别进行AR、PGK位点克隆性分析,所有标本均表现为相同的一条等位基因带完全消失(克隆率=0)或不均衡的甲基化模式(克隆率<0·25)。结论PSH组织中的多角形细胞和表面立方细胞具有相同的单克隆增生模式,提示二者可能均为肿瘤的实质细胞。

Objective To study the clonality of polygonal cells and surface cuboidal cells in the socalled pulmonary sclerosing hemangioma （PSH）. Methods 17 female surgically resected PSH were found. The polygonal cells and surface cuboidal cells of the 17 PSH cases were microdissected from routine hematoxylin and eosin-stained sections. Genomic DNA was extracted, pretreated through incubation with methylation-sensitive restrictive endonuclease HhaⅠ or HpaⅡ, and amplified by nested polymerase chain reaction for X chromosome-linked androgen receptor （AR） and phosphoglycerate kinase （PGK） genes. The length polymorphism of AR gene was demonstrated by denaturing polyacrylamide gel electrophoresis and silver staining. The PGK gone products were treated with Bst XI and resolved on agarose gel. Results Amongst the 17 female cases of PSH, 15 samples were successfully amplified for AR and PGK genes. The rates of polymorphism were 53% （8/15） and 27% （4/15） for AR and PGK genes respectively. Polygonal cells and surface cnboidal cells of 10 cases which were suitable for clonality study, showed the same loss of alleles （ clonality ratio = 0） or unbalanced methylation pattern （ clonality ratio 〈 0. 25 ）. Conclusion The polygonal cells and surface cnboidal cells in PSH demonstrate patterns of monoclonal proliferation, indicating that both represent true neoplastic cells.