de Jong Jeroen, Stoop Hans, Gillis Ad J M, van Gurp Ruud J H L M, van Drunen Ellen, Beverloo H Berna, Lau Yun-Fai Chris, Schneider Dominik T, Sherlock Jon K, Baeten John, Hatakeyama Shingo, Ohyama Chikara, Oosterhuis J Wolter, Looijenga Leendert H J
Department of Pathology, Erasmus MC-University Medical Center Rotterdam, Josephine Nefkens Institute, Daniel den Hoed Cancer Center.
Int J Androl. 2007 Aug;30(4):350-65. doi: 10.1111/j.1365-2605.2007.00802.x.
The JKT-1 cell line has been used in multiple independent studies as a representative model of human testicular seminoma. However, no cell line for this specific tumour type has been independently confirmed previously; and therefore, the seminomatous origin of JKT-1 must be proven. The genetic constitution of the JKT-1 cells was determined using flow cytometry and spectral karyotyping, as well as array comparative genomic hybridization and fluorescent in situ hybridization. Marker profiling, predominantly based on differentially expressed proteins during normal germ cell development, was performed by immunohistochemistry and Western blot analyses. Moreover, genome wide affymetrix mRNA expression and profiling of 157 microRNAs was performed, and the status of genomic imprinting was determined. A germ cell origin of the JKT-1 cells was in line with genomic imprinting status and marker profile (including positive staining for several cancer-testis antigens). However, the supposed primary tumour, from which the cell line was derived, being indeed a classical seminoma, was molecularly proven not to be the origin of the cell line. The characteristic chromosomal anomalies of seminoma, e.g. gain of the short arm of chromosome 12, as well as the informative marker profile (positive staining for OCT3/4, NANOG, among others) were absent in the various JKT-1 cell lines investigated, irrespective of where the cells were cultured. All results indicate that the JKT-1 cell line is not representative of human seminoma. Although it can originate from an early germ cell, a non-germ cell derivation cannot be excluded.
JKT-1细胞系已在多项独立研究中用作人类睾丸精原细胞瘤的代表性模型。然而,此前尚无针对这种特定肿瘤类型的细胞系得到独立确认;因此,必须证明JKT-1的精原细胞瘤起源。使用流式细胞术和光谱核型分析,以及阵列比较基因组杂交和荧光原位杂交来确定JKT-1细胞的基因构成。通过免疫组织化学和蛋白质印迹分析进行标志物分析,主要基于正常生殖细胞发育过程中差异表达的蛋白质。此外,还进行了全基因组Affymetrix mRNA表达和157种微小RNA的分析,并确定了基因组印记状态。JKT-1细胞的生殖细胞起源与基因组印记状态和标志物谱(包括几种癌-睾丸抗原的阳性染色)一致。然而,分子生物学证明,该细胞系所源自的假定原发性肿瘤实际上是经典精原细胞瘤,但并非该细胞系的起源。在所研究的各种JKT-1细胞系中,无论细胞在何处培养,均未出现精原细胞瘤的特征性染色体异常,例如12号染色体短臂的增加,以及信息丰富的标志物谱(OCT3/4、NANOG等的阳性染色)。所有结果表明,JKT-1细胞系不代表人类精原细胞瘤。尽管它可能起源于早期生殖细胞,但不能排除非生殖细胞来源。