Indraccolo Stefano, Tisato Veronica, Agata Simona, Moserle Lidia, Ferrari Silvia, Callegaro Monia, Persano Luca, Palma Maurizia Dalla, Scaini Maria Chiara, Esposito Giovanni, Fassina Ambrogio, Nicoletto Ornella, Plebani Mario, Chieco-Bianchi Luigi, Amadori Alberto, D'Andrea Emma, Montagna Marco
Istituto Oncologico Veneto, Padova, Italy.
Eur J Cancer. 2006 Jul;42(10):1475-83. doi: 10.1016/j.ejca.2006.01.057. Epub 2006 Jun 8.
The BRCA1 gene is responsible for a high number of hereditary breast and ovarian cancers that cluster in families with a strong genetic predisposition. Despite intense investigation, the accumulating findings on BRCA1 biological functions have not yet been translated into specific therapeutic approaches, also due to the lack of suitable experimental models. The purpose of this study was to establish and characterize cell cultures and xenografts from patients with BRCA1 -/- ovarian cancers. We derived two ovarian cancer cell lines, termed PD-OVCA1 and PD-OVCA2, both from patients previously treated with chemotherapy, that propagate in SCID mice as well as in vitro for a limited number of passages. Both cell lines expressed cytokeratins and the CA125 tumour marker. A detailed molecular characterization highlighted both constitutive and somatic genetic events that abrogate BRCA1 gene function. Both cell lines were shown to lose the wild type BRCA1 allele; intriguingly, these deletions were apparently accompanied by gain of one or more copies of the mutant alleles. Finally, a genomic profile of major chromosomal aberrations was obtained by the Multiplex Ligation-dependent Probe Amplification (MLPA) technique, which disclosed chromosomal imbalances targeting specific genes in each cell line. The PD-OVCA1 and PD-OVCA2 ovarian cancer cell lines will provide a valuable tool for new experimental models for the study of BRCA1-associated tumour biology.
BRCA1基因与大量遗传性乳腺癌和卵巢癌相关，这些癌症在具有强烈遗传易感性的家族中聚集。尽管进行了深入研究，但由于缺乏合适的实验模型，关于BRCA1生物学功能的累积发现尚未转化为具体的治疗方法。本研究的目的是建立并表征来自BRCA1基因敲除的卵巢癌患者的细胞培养物和异种移植模型。我们从两名先前接受过化疗的患者中获得了两种卵巢癌细胞系，分别命名为PD-OVCA1和PD-OVCA2，它们在SCID小鼠体内以及体外传代有限次数的情况下均可增殖。这两种细胞系均表达细胞角蛋白和CA125肿瘤标志物。详细的分子表征突出了消除BRCA1基因功能的组成性和体细胞遗传事件。两种细胞系均显示失去野生型BRCA1等位基因；有趣的是，这些缺失显然伴随着一个或多个突变等位基因拷贝的增加。最后，通过多重连接依赖探针扩增（MLPA）技术获得了主要染色体畸变的基因组图谱，该图谱揭示了每个细胞系中靶向特定基因的染色体失衡情况。PD-OVCA1和PD-OVCA2卵巢癌细胞系将为研究BRCA1相关肿瘤生物学的新实验模型提供有价值的工具。
