Detection of genomic imbalances in microdissected Hodgkin and Reed-Sternberg cells of classical Hodgkin's lymphoma by array-based comparative genomic hybridization
Detection of genomic imbalances in microdissected Hodgkin and Reed-Sternberg cells of classical Hodgkin's lymphoma by array-based comparative genomic hybridization
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DOI：
10.3324/haematol.12875
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发表时间：
2008-09-01
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影响因子：
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通讯作者：
Hansmann, Martin-Leo
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作者：
Hartmann, Sylvia;Martin-Subero, Jose I.;Hansmann, Martin-Leo
背景由于经典型霍奇金淋巴瘤组织中肿瘤性Hodgkin-Reed-斯滕贝格伯格细胞含量低，限制了其细胞遗传学分析。然而，现有的细胞遗传学数据指向极端的核型复杂性。为了深入了解经典霍奇金淋巴瘤的染色体不平衡，我们应用基于阵列的比较基因组杂交（阵列比较基因组杂交），使用来自显微切割的Hodgkin-Reed-斯滕贝格cells.Design and MethodsTo避免阵列比较基因组杂交的DNA扩增引入的偏差，选择了富含Hodgkin-Reed-斯滕贝格细胞的cHL病例。将从10例典型霍奇金淋巴瘤病例中每例的约100，000个显微切割的霍奇金-里德-斯滕贝格细胞获得的DNA杂交到商业105 K寡核苷酸比较基因组杂交微阵列上。通过间期细胞遗传学和定量聚合酶链反应分析证实了所选的不平衡，并在一个独立的经典霍奇金淋巴瘤系列中进一步研究。结果在至少五个cHL中发现的增益影响2 p12 -16，5 q15 -23，6p 22，8 q13，8 q24，9 p21 -24，9 q34，12 q13 -14，17 q12，19 p13，19 q13和20 q11，而至少5例病例中复发性丢失涉及Xp 21、6 q23 -24和13 q22。通过间期细胞遗传学和聚合酶链反应分析分别证实了所选基因的拷贝数变化和CDKN 2B（p15）基因的小缺失（156 kb）。获得的几个区域包括cHL中组成型表达的基因。其中，STAT 6（12 q13），NOTCH 1（9 q34）和JUNB（19 p13）的收益存在于额外的cHL通常低霍奇金-里德-斯滕贝格细胞content.ConclusionsThe本研究表明，阵列比较基因组杂交的显微切割霍奇金-里德-斯滕贝格细胞是适合于识别和表征染色体不平衡。受霍奇金-里德-斯坦伯格细胞中基因组变化影响的区域包括cHL中组成型表达的基因。
BackgroundCytogenetic analysis of classical Hodgkin's lymphoma is limited by the low content of the neoplastic Hodgkin-Reed-Sternberg cells in the affected tissues. However, available cytogenetic data point to an extreme karyotype complexity. To obtain insights into chromosomal imbalances in classical Hodgkin's lymphoma, we applied array-based comparative genomic hybridization (array comparative genomic hybridization) using DNA from microdissected Hodgkin-Reed-Sternberg cells.Design and MethodsTo avoid biases introduced by DNA amplification for array comparative genomic hybridization, cHL cases rich in Hodgkin-Reed-Sternberg cells were selected. DNA obtained from approximately 100,000 microdissected Hodgkin-Reed-Sternberg cells of each of ten classical Hodgkin's lymphoma cases was hybridized onto commercial 105 K oligonucleotide comparative genomic hybridization microarrays. Selected imbalances were confirmed by interphase cytogenetics and quantitative polymerase chain reaction analysis and further studied in an independent series of classical Hodgkin's lymphoma.ResultsGains identified in at least five cHL affected 2p12-16, 5q15-23, 6p22, 8q13, 8q24, 9p21-24, 9q34, 12q13-14, 17q12, 19p13, 19q13 and 20q11 whereas losses recurrent in at least five cases involved Xp21, 6q23-24 and 13q22. Copy number changes of selected genes and a small deletion (156 kb) of the CDKN2B (p15) gene were confirmed by interphase cytogenetics and polymerase chain reaction analysis, respectively. Several gained regions included genes constitutively expressed in cHL. Among these, gains of STAT6 (12q13), NOTCH1 (9q34) and JUNB (19p13) were present in additional cHL with the usual low Hodgkin-Reed-Sternberg cell content.ConclusionsThe present study demonstrates that array comparative genomic hybridization of microdissected Hodgkin-Reed-Sternberg cells is suitable for identifying and characterizing chromosomal imbalances. Regions affected by genomic changes in Hodgkin-Reed-Stemberg cells recurrently include genes constitutively expressed in cHL.