Subclones in B-lymphoma cell lines: isogenic models for the study of gene regulation.
Subclones in B-lymphoma cell lines: isogenic models for the study of gene regulation.
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DOI：
10.18632/oncotarget.11524
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发表时间：
2016-09-27
期刊：
影响因子：
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通讯作者：
Drexler HG
中科院分区：
文献类型：
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作者：
Quentmeier H;Pommerenke C;Ammerpohl O;Geffers R;Hauer V;MacLeod RA;Nagel S;Romani J;Rosati E;Rosén A;Uphoff CC;Zaborski M;Drexler HG
遗传异质性虽然在肿瘤中很常见，但在细胞系中很少有记录。为了检查B淋巴瘤细胞系由亚克隆组成的频率，我们进行了免疫球蛋白（IG）重链超突变分析。揭示了亚克隆在B细胞淋巴瘤细胞系中并不罕见，6/49个IG超突变细胞系（12%）由具有单个IG突变的亚克隆组成。还在2/284个表现出双峰CD标记物表达的白血病/淋巴瘤细胞系中鉴定了亚克隆。我们成功地从四个细胞系（HG 3，SU-DHL-5，TMD-8，U-2932）中分离出10个亚克隆。进行全外显子组测序以从分子上表征这些亚克隆。我们详细描述了细胞系HG 3的克隆结构，来自慢性淋巴细胞白血病。HG 3由三个亚克隆组成，每个亚克隆具有克隆特异性畸变、基因表达和DNA甲基化模式。虽然供体患者白血病细胞是CD 5+，但三个HG 3亚克隆中的两个独立地丢失了该标记。HG 3细胞上的CD 5受表观遗传/转录机制调控，而不是迄今报道的选择性剪接。总之，我们表明，在携带个别突变的细胞系中存在亚克隆，并以差异表达基因集为特征并不罕见。我们还表明，这些亚克隆可以是有用的调控和功能研究的同基因模型。
Genetic heterogeneity though common in tumors has been rarely documented in cell lines. To examine how often B-lymphoma cell lines are comprised of subclones, we performed immunoglobulin (IG) heavy chain hypermutation analysis. Revealing that subclones are not rare in B-cell lymphoma cell lines, 6/49 IG hypermutated cell lines (12%) consisted of subclones with individual IG mutations. Subclones were also identified in 2/284 leukemia/lymphoma cell lines exhibiting bimodal CD marker expression. We successfully isolated 10 subclones from four cell lines (HG3, SU-DHL-5, TMD-8, U-2932). Whole exome sequencing was performed to molecularly characterize these subclones. We describe in detail the clonal structure of cell line HG3, derived from chronic lymphocytic leukemia. HG3 consists of three subclones each bearing clone-specific aberrations, gene expression and DNA methylation patterns. While donor patient leukemic cells were CD5+, two of three HG3 subclones had independently lost this marker. CD5 on HG3 cells was regulated by epigenetic/transcriptional mechanisms rather than by alternative splicing as reported hitherto. In conclusion, we show that the presence of subclones in cell lines carrying individual mutations and characterized by sets of differentially expressed genes is not uncommon. We show also that these subclones can be useful isogenic models for regulatory and functional studies.