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人卵巢癌细胞系CABA I:一种独特的基因进化

The human ovarian cancer cell line CABA I: A peculiar genetic evolution.

作者信息

Giusti Ilaria, Cervelli Carla, D'Ascenzo Sandra, Di Francesco Marianna, Ligas Claudio, D'Alessandro Elvira, Papola Franco, Dolo Vincenza

机构信息

Department of Life, Health and Environmental Sciences, University of L'Aquila, L'Aquila, Italy.

Regional Center of Immunohematology and Tissue Typing, L'Aquila, Italy.

出版信息

Int J Mol Med. 2016 Apr;37(4):879-88. doi: 10.3892/ijmm.2016.2501. Epub 2016 Feb 25.

Abstract

The objective of this study was to study the human ovarian cancer cell line CABA I by means of short tandem repeats (STR) profiling and cytogenetic analysis in order to prevent future misidentification or cross-contamination and verify its stability during in vitro cultivation. To this end, cells at passages 18 and 38 were analyzed using cytogenetic techniques in order to verify possible chromosomal aberrations and the karyotypic evolution of this cell line; GTG-banding and FISH were also performed. For STR analysis, DNA was extracted using the automated extractor MagNA pure and analyzed by means of PowerPlex 16 HS. STR profiles were analyzed by GeneMapper 3.2.1 software. Whereas comparative cytogenetic analysis of CABA I cells at passage 18 and 38 has demonstrated considerable genetic instability, we found that STR profiles were essentially unaltered in both analyzed passages, suggesting that the STR profile is reliable and could be used for the regular authentication of CABA I over time. It should be emphasized, however, that of the 16 loci generally used in human STR profiles, only 3 were properly detectable in CABA I. The data highlight that the CABA I cell line demonstrates an anomalous STR profile that does not fully adjust the criteria currently used for the identification of human cells; in spite of this, it remains stable during the in vitro maintainance. Moreover, the genetic instability of the CABA I cell line overlaps with those observed in vivo in tumor cells, making it a suitable candidate to analyze, in vitro, the peculiar genetic evolution of ovarian cancer cells.

摘要

本研究的目的是通过短串联重复序列(STR)分析和细胞遗传学分析来研究人卵巢癌细胞系CABA I,以防止未来的错误鉴定或交叉污染,并验证其在体外培养过程中的稳定性。为此,使用细胞遗传学技术对第18代和第38代细胞进行分析,以验证该细胞系可能存在的染色体畸变和核型演变;还进行了GTG显带和荧光原位杂交(FISH)。对于STR分析,使用自动提取仪MagNA pure提取DNA,并通过PowerPlex 16 HS进行分析。STR图谱通过GeneMapper 3.2.1软件进行分析。虽然对第18代和第38代CABA I细胞的比较细胞遗传学分析表明存在相当大的遗传不稳定,但我们发现两个分析代次的STR图谱基本未改变,这表明STR图谱是可靠的,可用于随时间对CABA I进行定期鉴定。然而,应该强调的是,在人类STR图谱中通常使用的16个位点中,在CABA I中只有3个能够被正确检测到。数据突出表明,CABA I细胞系显示出异常的STR图谱,不完全符合目前用于鉴定人类细胞的标准;尽管如此,它在体外培养过程中仍保持稳定。此外,CABA I细胞系的遗传不稳定与肿瘤细胞在体内观察到的情况重叠,使其成为在体外分析卵巢癌细胞特殊遗传演变的合适候选者。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7fcc/4790663/44de24020b23/IJMM-37-04-0879-g00.jpg

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