Glaser R, Zhang H Y, Yao K T, Zhu H C, Wang F X, Li G Y, Wen D S, Li Y P
Department of Medical Microbiology and Immunology, Ohio State University Medical Center, Columbus 43210.
Proc Natl Acad Sci U S A. 1989 Dec;86(23):9524-8. doi: 10.1073/pnas.86.23.9524.
Two epithelial tumor cell lines were established from biopsy specimens of nasopharyngeal carcinomas (NPC). The specimens were taken from poorly differentiated squamous cell carcinomas of the nasopharynx. The tissues were prepared for cell culture and eventually two continuous epithelial cell lines were obtained and designated HONE-1 and HNE-1. Light and electron microscopic examination of these two cell lines demonstrated cells with an epithelial morphology including the presence of desmosomes. The HNE-1 cell line has been passaged more than 100 times and the HONE-1 cell line has been passaged more than 90 times. It was found that early-passage uncloned HNE-1 cells (passage 23) could be superinfected with the B95-8 and NPC-EBV isolates as demonstrated by the induction of Epstein-Barr virus (EBV)-specific early antigen(s) in a small percentage of the cells; HONE-1 cells could also be superinfected with EBV. Southern blot analysis detected EBV DNA in samples from uncloned HNE-1 cells at passages 12, 17, 21, 27, and 35. However, by passage 45, EBV DNA could no longer be detected in HNE-1 cells by Southern blot analysis. The EBV genome was detected in parental HONE-1 cells at subculture 9 and in clone 40 cells up to passage 40 thus far. When HNE-1 cells were examined for the expression of the EBV-encoded nuclear antigen (EBNA) at passage 12, only about 10% of the cells were found to be positive. The percentage of EBNA-positive HNE-1 cells decreased as the cells were passaged. A similar loss of EBNA was observed in uncloned HONE-1 cells, but not in HONE-1 clone 40 cells. In clone 40, which has been passaged 40 times thus far, 85-90% of the cells are still EBNA-positive. The data suggest that EBV genome-positive HNE-1 and HONE-1 cells were lost as the cells were cultivated in vitro and that cloning the cells at an early passage level may be critical in maintaining EBV genome-positive epithelial NPC cells. These EBV genome-positive epithelial NPC cell lines will be useful for studying the association of EBV and NPC.
从鼻咽癌（NPC）活检标本中建立了两种上皮肿瘤细胞系。标本取自鼻咽低分化鳞状细胞癌。将组织制备用于细胞培养，最终获得了两种连续的上皮细胞系，命名为HONE - 1和HNE - 1。对这两种细胞系进行光镜和电镜检查，结果显示细胞具有上皮形态，包括存在桥粒。HNE - 1细胞系已传代100多次，HONE - 1细胞系已传代90多次。研究发现，早期传代未克隆的HNE - 1细胞（第23代）可被B95 - 8和NPC - EBV分离株超感染，少数细胞中可诱导出爱泼斯坦 - 巴尔病毒（EBV）特异性早期抗原；HONE - 1细胞也可被EBV超感染。Southern印迹分析在第12、17、21、27和35代未克隆的HNE - 1细胞样本中检测到EBV DNA。然而，到第45代时，通过Southern印迹分析在HNE - 1细胞中已无法检测到EBV DNA。在原代HONE - 1细胞传代9时以及克隆40细胞传代至第40代时均检测到EBV基因组。当在第12代检测HNE - 1细胞中EBV编码的核抗原（EBNA）表达时，仅发现约10%的细胞呈阳性。随着细胞传代，EBNA阳性的HNE - 1细胞百分比下降。在未克隆的HONE - 1细胞中也观察到类似的EBNA丢失情况，但在HONE - clone 40细胞中未观察到。在已传代40次的克隆40中，85 - 90%的细胞仍为EBNA阳性。数据表明，EBV基因组阳性的HNE - 1和HONE - 1细胞在体外培养过程中丢失，并且在早期传代水平对细胞进行克隆可能对维持EBV基因组阳性的上皮性NPC细胞至关重要。这些EBV基因组阳性的上皮性NPC细胞系将有助于研究EBV与NPC的关联。
