Institute for Research on Cancer and Aging, University of Nice-Sophia-Antipolis, Nice, France.
Virchows Arch. 2012 Jul;461(1):67-78. doi: 10.1007/s00428-012-1256-5. Epub 2012 Jun 8.
While surgery is the usual treatment for localized well-differentiated and dedifferentiated liposarcomas (WDLPS/DDLPS), the therapeutic options for patients with advanced disease are limited. The classical antimitotic treatments are most often inefficient. The establishment of genetically characterized cell lines is therefore crucial for providing in vitro models for novel targeted therapies. We have used spectral karyotyping, fluorescence in situ hybridization with whole chromosome painting and locus-specific probes, and array-comparative genomic hybridization to identify the chromosomal and molecular alterations of a novel cell line established from a recurring sclerosing WDLPS. The karyotype was hypertriploid and showed multiple structural anomalies. All cells retained the presence of a giant marker chromosome that had been previously identified in the primary cell cultures. This giant chromosome contained high-level amplification of chromosomal regions 12q13-21 and lacked the alpha-satellite centromeric sequences associated with WDLPS/DDLPS. The 12q amplicon was large, containing 370 amplified genes. The DNA copy number ranged from 3 to 57. The highest levels of amplification were observed at 12q14.3 for GNS, WIF1, and HMGA2. We analyzed the mRNA expression status by real-time reverse transcription polymerase chain reaction for six genes from this amplicon: MDM2, HMGA2, CDK4, TSPAN31, WIF1, and YEATS4. mRNA overexpression was correlated with genomic amplification. A second amplicon originating from 10p11-14 was also present in the giant marker chromosome. The 10p amplicon contained 62 genes, including oncogenes such as MLLT10, previously described in chimeric fusion with MLL in leukemias, NEBL, and BMI1.
虽然手术是治疗局限性高分化和去分化脂肪肉瘤(WDLPS/DDLPS)的常用方法,但晚期患者的治疗选择有限。经典的抗有丝分裂治疗通常无效。因此,建立具有遗传特征的细胞系对于提供新的靶向治疗的体外模型至关重要。我们使用光谱核型分析、全染色体涂染和基因特异性探针的荧光原位杂交以及阵列比较基因组杂交来鉴定从复发性硬化性 WDLPS 中建立的新型细胞系的染色体和分子改变。核型为超三倍体,并显示出多种结构异常。所有细胞均保留了先前在原代细胞培养中鉴定出的巨型标记染色体。该巨型染色体包含染色体区域 12q13-21 的高水平扩增,并且缺乏与 WDLPS/DDLPS 相关的α卫星着丝粒序列。12q 扩增子很大,包含 370 个扩增基因。DNA 拷贝数范围为 3 至 57。在 12q14.3 处观察到最高水平的扩增,用于 GNS、WIF1 和 HMGA2。我们通过实时逆转录聚合酶链反应分析来自该扩增子的六个基因的 mRNA 表达状态:MDM2、HMGA2、CDK4、TSPAN31、WIF1 和 YEATS4。mRNA 过表达与基因组扩增相关。第二个源自 10p11-14 的扩增子也存在于巨型标记染色体中。10p 扩增子包含 62 个基因,包括先前在白血病中与 MLL 融合的嵌合融合描述的癌基因,如 MLLT10、NEBL 和 BMI1。