Sery T W, Lee E Y, Lee W H, Bookstein R, Wong V, Shields J A, Augsburger J J, Donoso L A
Research Department, Wills Eye Hospital, Philadelphia, Pa. 19107.
J Pediatr Ophthalmol Strabismus. 1990 Jul-Aug;27(4):212-7. doi: 10.3928/0191-3913-19900701-11.
From 49 eyes enucleated for retinoblastoma, two new cell lines, WERI-Rb24 (W-24) and WERI-Rb27 (W-27), were established in long-term culture (greater than 5 years). The W-24 cell line was derived from a 22-month-old boy with sporadic retinoblastoma; the W-27 cell line was derived from a 24-month-old boy with bilateral retinoblastoma. Both cell lines show abnormal mRNA transcripts corresponding to the retinoblastoma gene. At the DNA level, one retinoblastoma allele was not present in the W-24 cell line. In the W-27 cell line a mutation was identified in one allele, while the other appears grossly normal. Since no normal retinoblastoma mRNA can be detected in either cell line, a subtle mutation must occur on the grossly normal allele. Comparison of DNA in W-27 lymphocytes and tumor cells with DNA probe p6NR-0.5 indicates that the observed mutation occurred somatically. The application of these two new retinoblastoma cell lines to the characterization of defects in the retinoblastoma gene and to gene replacement therapy in retinoblastoma is discussed.
从49只因视网膜母细胞瘤而摘除的眼球中，建立了两个长期培养（超过5年）的新细胞系，即WERI-Rb24（W-24）和WERI-Rb27（W-27）。W-24细胞系源自一名患有散发性视网膜母细胞瘤的22个月大男孩；W-27细胞系源自一名患有双侧视网膜母细胞瘤的24个月大男孩。这两个细胞系均显示出与视网膜母细胞瘤基因相对应的异常mRNA转录本。在DNA水平上，W-24细胞系中不存在一个视网膜母细胞瘤等位基因。在W-27细胞系中，一个等位基因被鉴定出发生了突变，而另一个看起来大体正常。由于在这两个细胞系中均未检测到正常的视网膜母细胞瘤mRNA，因此在大体正常的等位基因上必定发生了微小突变。用DNA探针p6NR-0.5对W-27淋巴细胞和肿瘤细胞中的DNA进行比较，表明观察到的突变是体细胞发生的。讨论了这两个新的视网膜母细胞瘤细胞系在视网膜母细胞瘤基因缺陷特征分析和基因替代治疗中的应用。
