Division of Haematology/Oncology, CHUS University Hospital, 3001 12th Avenue North, Sherbrooke, Québec, Canada.
Lab Invest. 2010 Apr;90(4):611-9. doi: 10.1038/labinvest.2010.2. Epub 2010 Feb 8.
In Epstein-Barr virus (EBV) negative Hodgkin's cell lines and classical EBV-negative Hodgkin's lymphoma (HL), Reed-Sternberg cells (RS cells) represent end-stage tumor cells, in which further nuclear division becomes impossible because of sustained telomere loss, shortening and aggregation. However, the three-dimensional (3D) telomere organization in latent membrane protein 1 (LMP1)-expressing RS cells of EBV-associated HL is not known. We performed a 3D telomere analysis after quantitative fluorescent in situ hybridization on 5 mum tissue sections on two LMP1-expressing HL cases and showed highly significant telomere shortening (P<0.0001) and formation of telomere aggregates in RS cells (P<0.0001), when compared with the mononuclear precursor Hodgkin cells (H cells). Telomere-poor or telomere-free 'ghost' nuclei were a regular finding in these RS cells. These nuclei and their telomere content strongly contrasted with the corona of surrounding lymphocytes showing numerous midsized telomere hybridization signals. Both H cells and RS cells of two EBV-negative HL cases analyzed in parallel showed 3D telomere patterns identical to those of LMP1-expressing cases. As a major advance, our 3D nuclear imaging approach allows the visualization of hitherto unknown profound changes in the 3D nuclear telomere organization associated with the transition from LMP1-positive H cells to LMP1-positive RS cells. We conclude that RS cells irrespective of LMP1 expression are end-stage tumor cells in which the extent of their inability to divide further is proportional to the increase of very short telomeres, telomere loss, aggregate formation and the generation of 'ghost' nuclei.
在 Epstein-Barr 病毒（EBV）阴性霍奇金细胞系和经典 EBV 阴性霍奇金淋巴瘤（HL）中，Reed-Sternberg 细胞（RS 细胞）代表终末肿瘤细胞，由于持续的端粒丢失、缩短和聚集，进一步的核分裂变得不可能。然而，潜伏膜蛋白 1（LMP1）表达的 EBV 相关 HL 的 RS 细胞中的三维（3D）端粒组织尚不清楚。我们对两个 LMP1 表达的 HL 病例的 5 µm 组织切片进行了定量荧光原位杂交 3D 端粒分析，并显示与单核前体霍奇金细胞（H 细胞）相比，RS 细胞中端粒显著缩短（P<0.0001）和端粒聚集（P<0.0001）。在这些 RS 细胞中，经常可以发现端粒贫乏或无端粒的“幽灵”核。这些核及其端粒含量与周围淋巴细胞的冠形成鲜明对比，显示出大量中等大小的端粒杂交信号。同时分析的两个 EBV 阴性 HL 病例的 H 细胞和 RS 细胞显示出与 LMP1 表达病例相同的 3D 端粒模式。作为一个主要的进步，我们的 3D 核成像方法允许可视化与从 LMP1 阳性 H 细胞到 LMP1 阳性 RS 细胞的转变相关的 3D 核端粒组织中未知的深刻变化。我们得出结论，无论是否表达 LMP1，RS 细胞都是终末肿瘤细胞，其进一步分裂能力的丧失程度与极短端粒、端粒丢失、聚集形成和“幽灵”核的产生成正比。
