Department of Medicine, Division of Medical Oncology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.
Gynecol Oncol. 2012 Oct;127(1):241-8. doi: 10.1016/j.ygyno.2012.06.017. Epub 2012 Jun 16.
Cell lines derived from human ovarian and endometrial cancers, and their immortalized non-malignant counterparts, are critical tools to investigate and characterize molecular mechanisms underlying gynecologic tumorigenesis, and facilitate development of novel therapeutics. To determine the extent of misidentification, contamination and redundancy, with evident consequences for the validity of research based upon these models, we undertook a systematic analysis and cataloging of endometrial and ovarian cell lines.
Profiling of cell lines by analysis of DNA microsatellite short tandem repeats (STR), p53 nucleotide polymorphisms and microsatellite instability was performed.
Fifty-one ovarian cancer lines were profiled with ten found to be redundant and five (A2008, OV2008, C13, SK-OV-4 and SK-OV-6) identified as cervical cancer cells. Ten endometrial cell lines were analyzed, with RL-92, HEC-1A, HEC-1B, HEC-50, KLE, and AN3CA all exhibiting unique, uncontaminated STR profiles. Multiple variants of Ishikawa and ECC-1 endometrial cancer cell lines were genotyped and analyzed by sequencing of mutations in the p53 gene. The profile of ECC-1 cells did not match the EnCa-101 tumor, from which it was reportedly derived, and all ECC-1 isolates were genotyped as Ishikawa cells, MCF-7 breast cancer cells, or a combination thereof. Two normal, immortalized endometrial epithelial cell lines, HES cells and the hTERT-EEC line, were identified as HeLa cervical carcinoma and MCF-7 breast cancer cells, respectively.
Results demonstrate significant misidentification, duplication, and loss of integrity of endometrial and ovarian cancer cell lines. Authentication by STR DNA profiling is a simple and economical method to verify and validate studies undertaken with these models.
源自人类卵巢癌和子宫内膜癌的细胞系及其永生化的非恶性对应物，是研究和描述妇科肿瘤发生分子机制的重要工具，并有助于开发新的治疗方法。为了确定这些模型所基于的研究的有效性受到明显影响的错误识别、污染和冗余的程度，我们对子宫内膜和卵巢细胞系进行了系统的分析和编目。
通过分析 DNA 微卫星短串联重复序列（STR）、p53 核苷酸多态性和微卫星不稳定性对细胞系进行分析。
对 51 种卵巢癌细胞系进行了分析，其中 10 种被认为是冗余的，5 种（A2008、OV2008、C13、SK-OV-4 和 SK-OV-6）被鉴定为宫颈癌细胞。对 10 种子宫内膜细胞系进行了分析，其中 RL-92、HEC-1A、HEC-1B、HEC-50、KLE 和 AN3CA 均具有独特、无污染的 STR 谱。对多个 Ishikawa 和 ECC-1 子宫内膜癌细胞系进行了基因分型，并通过 p53 基因突变的测序对其进行了分析。ECC-1 细胞的特征与据报道源自该细胞的 EnCa-101 肿瘤并不匹配，并且所有 ECC-1 分离株均被鉴定为 Ishikawa 细胞、MCF-7 乳腺癌细胞或两者的组合。两种正常的、永生化的子宫内膜上皮细胞系 HES 细胞和 hTERT-EEC 系分别被鉴定为 HeLa 宫颈癌和 MCF-7 乳腺癌细胞。
结果表明子宫内膜癌和卵巢癌细胞系存在显著的错误识别、重复和完整性丧失。通过 STR DNA 分析进行鉴定是验证和验证这些模型所进行研究的简单而经济的方法。
