Use of microdissection and molecular genetics in the pathologic diagnosis of retinoblastoma.
Use of microdissection and molecular genetics in the pathologic diagnosis of retinoblastoma.
复制标题
显微切割和分子遗传学在视网膜母细胞瘤病理诊断中的应用。
DOI：
10.1097/00006982-199907000-00009
复制
发表时间：
1999
期刊：
影响因子：
--
通讯作者：
C. Chan
中科院分区：
文献类型：
--
作者：
S. Whitcup;W. Park;A. Gasch;R. Eagle;A. Filie;R. Nussenblatt;Z. Zhuang;C. Chan
背景/目的
视网膜母细胞瘤由视网膜母细胞瘤基因的两个等位基因的突变或缺失引起。虽然视网膜母细胞瘤通常在临床上被识别，但某些形式的疾病可能无法诊断。本研究的目的是确定是否使用分子遗传学检测杂合性丢失（洛）在视网膜母细胞瘤基因可以帮助眼科病理学家在这种恶性肿瘤的诊断。
方法
脱氧核糖核酸（DNA）是从两名弥漫性视网膜母细胞瘤患者的三个眼部标本显微解剖的肿瘤细胞中获得的。应用聚合酶链反应（PCR）检测视网膜母细胞瘤基因的两个微卫星标记（D13 S153和D13 S118）。当两个多态性等位基因中的一个存在于正常组织的DNA中，但在从肿瘤细胞获得的DNA中不存在或减少时，鉴定为杂合性丢失。
结果
两例弥漫性视网膜母细胞瘤患者的三份标本均存在杂合性缺失。在一名患者中，视网膜母细胞瘤的诊断是基于从玻璃体切除术获得的肿瘤细胞中鉴定出洛缺失。
结论
这项研究表明，洛鉴定视网膜母细胞瘤细胞不仅可以帮助我们了解这种肿瘤的分子遗传学，而且可以帮助眼科病理学家在诊断的非典型形式的疾病。
BACKGROUND/PURPOSE
Retinoblastoma results from mutations or loss of both alleles of the retinoblastoma gene. Although retinoblastoma is usually recognized clinically, some forms of the disease can elude diagnosis. The purpose of this study was to determine whether the use of molecular genetics to detect a loss of heterozygosity (LOH) in the retinoblastoma gene could assist the ocular pathologist in the diagnosis of this malignancy.
METHODS
Deoxyribonucleic acid (DNA) was obtained from tumor cells microdissected from three ocular specimens from two patients with diffuse retinoblastoma. Polymerase chain reaction was used to detect two microsatellite markers (D13S153 and D13S118) of the retinoblastoma gene. Loss of heterozygosity was identified when one of the two polymorphic alleles was present in the DNA from normal tissue but absent or reduced in the DNA obtained from tumor cells.
RESULTS
Loss of heterozygosity was identified in all three specimens from the two patients with diffuse retinoblastoma. In one patient, the diagnosis of retinoblastoma was based on identification of LOH from tumor cells obtained from vitrectomy.
CONCLUSIONS
This study demonstrates that identification of LOH in retinoblastoma cells not only can contribute to our understanding of the molecular genetics of this tumor, but also can help the ocular pathologist in the diagnosis of atypical forms of the disease.