Institute of Pathology, University Medical Center Duesseldorf, Duesseldorf, Germany.
Genes Chromosomes Cancer. 2010 Jan;49(1):40-51. doi: 10.1002/gcc.20717.
Usage of cancer cell lines has repeatedly generated conflicting results provoked by differences among subclones or contamination with mycoplasm or other immortal mammalian cells. To overcome these limitations, we decided within the EuroBoNeT consortium to characterize a common set of cell lines including osteosarcomas (OS), Ewing sarcomas (ES), and chondrosarcomas (CS). DNA fingerprinting was used to guarantee the identity of all of the cell lines and to distinguish subclones of osteosarcoma cell line HOS. Screening for homozygous loss of 38 tumor suppressor genes by MLPA revealed deletion of CDKN2A as the most common event (15/36), strictly associated with absence of the CDKN2A (p16) protein. Ten cell lines showed missense mutations of the TP53 gene while another set of nine cell lines showed mutations resulting in truncation of the TP53 protein. Cells harboring missense mutations expressed high levels of nuclear TP53, while cell lines with nonsense mutations showed weak/absent staining for TP53. TP53(wt) cell lines usually expressed the protein in 2-10% of the cells. However, seven TP53(wt) osteosarcomas were negative for both mRNA and protein expression. Our analyses shed light on the correlation between immunohistochemical and genetic data for CDKN2A and TP53, and confirm the importance of these signaling pathways. The characterization of a substantial number of cell lines represents an important step to supply research groups with proven models for further advanced studies on tumor biology and may help to make results from different laboratories more comparable.
癌细胞系的使用反复产生了冲突的结果，这些结果是由于亚克隆之间的差异或支原体或其他永生化哺乳动物细胞的污染引起的。为了克服这些限制，我们在 EuroBoNeT 联盟内决定对一组包括骨肉瘤 (OS)、尤文肉瘤 (ES) 和软骨肉瘤 (CS) 的常见细胞系进行特征描述。DNA 指纹分析用于保证所有细胞系的身份，并区分骨肉瘤细胞系 HOS 的亚克隆。通过 MLPA 对 38 个肿瘤抑制基因的纯合缺失进行筛选，发现 CDKN2A 的缺失是最常见的事件 (15/36)，与 CDKN2A(p16)蛋白的缺失严格相关。10 个细胞系显示 TP53 基因的错义突变，而另一组 9 个细胞系显示导致 TP53 蛋白截断的突变。携带错义突变的细胞表达高水平的核 TP53，而具有无意义突变的细胞系对 TP53 的染色较弱/缺失。携带错义突变的 TP53(wt)细胞系通常在 2-10%的细胞中表达该蛋白。然而，7 个 TP53(wt)骨肉瘤在 mRNA 和蛋白表达上均为阴性。我们的分析阐明了 CDKN2A 和 TP53 的免疫组织化学和遗传数据之间的相关性，并证实了这些信号通路的重要性。大量细胞系的特征描述是为研究小组提供经过验证的模型以进行肿瘤生物学的进一步深入研究的重要步骤，并且可能有助于使不同实验室的结果更加可比。
