Fu Jiewen, Cheng Jingliang, Liu Xiaoyan, Li Jun, Wei Chunli, Zheng Xiaoli, He Tao, Fu Junjiang
Key Laboratory of Epigenetics and Oncology, The Research Center for Preclinical Medicine, Southwest Medical University, Luzhou, 646000, Sichuan, China.
Judicial Authentication Center, Southwest Medical University, Luzhou, 646000, Sichuan, China.
Mol Biol Rep. 2018 Dec;45(6):2689-2695. doi: 10.1007/s11033-018-4438-7. Epub 2018 Nov 2.
Cancer cell lines are used worldwide in biomedical researches, and data interpretation solely depends on unambiguous attribution of those respective cell lines to its original sources. Approximately one-third of all cell lines have an origin other than that assumed, leading to invalid results. It is necessary to characterize the origin of cell lines. Short-tandem-repeat (STR) fingerprinting (DNA fingerprinting) is the method for characterization of genetic identity in cultured cell lines under certain experimental conditions. We showed the fingerprinting profiles in a summed and unidentified human cancer cell line comparison to HCC1954 cell line, revealing marked alterations in DNA fingerprinting profiles up to fourteen STR loci from 16 loci. Furthermore, Sanger DNA sequencing showed no c.3140A > G heterozygous mutation in the PIK3CA gene of this suspected HCC1954 cell line. In addition, we showed the fingerprinting profiles in an unidentified cancer cell line comparison to SiHa cervical cell line, revealing same DNA fingerprinting profiles. In conclusion, we have successfully authenticated and identified both suspected HCC1954 and SiHa cell lines by STR analysis and DNA sequencing. STR analysis combined DNA sequencing may be very useful to evaluate genotypes of cancer cell lines in our cancer studies, as well as in judicial authentication and forensic sciences.
癌细胞系在全球生物医学研究中被广泛使用,数据解读完全依赖于将这些各自的细胞系明确归属于其原始来源。所有细胞系中约有三分之一的起源并非如假设的那样,这导致结果无效。有必要对细胞系的起源进行表征。短串联重复序列(STR)指纹图谱(DNA指纹图谱)是在特定实验条件下对培养细胞系进行遗传身份表征的方法。我们展示了一个汇总且身份不明的人类癌细胞系与HCC1954细胞系比较的指纹图谱,揭示了在16个位点中多达14个STR位点的DNA指纹图谱存在显著改变。此外,桑格DNA测序显示这个疑似HCC1954细胞系的PIK3CA基因中不存在c.3140A > G杂合突变。另外,我们展示了一个身份不明的癌细胞系与SiHa宫颈癌细胞系比较的指纹图谱,揭示了相同的DNA指纹图谱。总之,我们通过STR分析和DNA测序成功鉴定并确认了疑似HCC1954和SiHa细胞系。STR分析结合DNA测序在我们的癌症研究以及司法鉴定和法医学中,对于评估癌细胞系的基因型可能非常有用。