Flow cytometric sorting of paraffin-embedded tumor tissues considerably improves molecular genetic analysis

被引:16
|
作者
Jordanova, ES
Corver, WE
Vonk, MJ
Leers, MPG
Riemersma, SA
Schuuring, E
Kluin, PM
机构
[1] Leiden Univ, Ctr Med, Dept Pathol, NL-2333 ZA Leiden, Netherlands
[2] Atrium Med Ctr Heerlen, Heerlen, Netherlands
[3] Univ Groningen Hosp, Groningen, Netherlands
关键词
paraffin-embedded tissue; flow cytometric sorting; B-cell lymphoma; HLA; loss of heterozygosity; LOH; flow cytometry; fluorescent in situ hybridization; FISH;
D O I
10.1309/HPR11R7LQ9NNCCG8
中图分类号
R36 [病理学];
学科分类号
100104 ;
摘要
The characterization of genetic aberrations in paraffin-embedded tumor material is impaired by contaminating normal cells. In the present study on the genetic causes of loss of HLA expression in diffuse large B-cell lymphoma (DLBCL), we compared the efficacy of microdissection with flow cytometric sorting of tumor cells. Single-cell suspensions from paraffin-embedded material of 5 DLBCL cases were stained for CD79a and DNA content (propidium iodide). Fluorescent in situ hybridization (FISH) using HLA class II and chromosome 6 centromeric probes and loss of heterozygosity (LOH) analysis with 5 HLA-specific microsatellite markers were performed on microdissected and flow cytometry-sorted fractions. FISH confirmed considerable enrichment of the samples after flow cytometric sorting and disclosed tumor heterogeneity in 4 cases. Moreover, lymphomas with a so-called zebra LOH pattern in the microdissected material showed unambiguous LOH after flow cytometric sorting, revealing in 1 case a biologically relevant hemizygous deletion in the HLA region.
引用
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页码:327 / 334
页数:8
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