Department of Pathology, University of Valencia, Valencia, Spain.
Diagn Pathol. 2008 Jul 15;3 Suppl 1(Suppl 1):S27. doi: 10.1186/1746-1596-3-S1-S27.
Tissue microarrays (TMAs) are used to study genomics and proteomics in several tumour tissue samples. Cell lines (CC) are of great importance in the study of the genetic changes in tumours, and some reveal several aspects of tumour oncogenesis. There are few published reports on Ewing's tumours with TMAs including original tumours (OT) and corresponding CC.
We have performed four TMAs, from 3 OT and the corresponding CC of successive in vivo and in vitro tumour passages. Xenotransplant CC in nude mice from OT (XT/OT) was made. Subsequently multiple XT were performed and in vitro XT cell line (CC/XT) was obtained. In vivo re-inoculation of CC/XT (XT/CC) was planned. TMAs with the successive tumour passages that grew in nude mice (XT/OT and XT/CC) were analyzed by morphologic pattern (Hematoxilin/eosin), immunohistochemical staining (CD99, FLI1, p16, p53, ki-67), fluorescent in situ hybridization-FISH-(EWSR1 break apart, p16 and p53 status) and gene fusion types.
Heterogeneous results of the p16, p53 and ki67 in OT, XT/OT, CC/XT and XT/CC were observed. The three cell lines revealed EWS/FLI1 rearrangements. p16 gene was deleted only in one case. The deletion was detected by FISH and confirmed by PCR assays. A p53 alteration was found in the second case with monosomy and subsequently polysomic status of chromosome 17 during the evolution of CC. The PCR study revealed p53 mutation. The third case showed hypermethylation in the promoter of p16. The growth of the tumour in nude mice was more accelerated when the inoculation was performed from the CC/XT, increasing progressively over the passages. The third case did not reveal tumour growth in nude mice after the re-inoculation of CC/XT.
The study of several cores from original tumours and successive tumour passages in TMAs facilitated the analysis of the genetic alteration and protein expression in Ewing's tumours.
组织微阵列（TMA）用于研究多个肿瘤组织样本的基因组学和蛋白质组学。细胞系（CC）在肿瘤遗传变化的研究中非常重要，有些揭示了肿瘤发生的多个方面。关于包含原始肿瘤（OT）和相应 CC 的 Ewing 肿瘤的 TMA 报道很少。
我们已经进行了四个 TMA，来自 3 个 OT 和连续的体内和体外肿瘤传代的相应 CC。从 OT 进行了异种移植 CC 在裸鼠中的移植（XT/OT）。随后进行了多次 XT，并获得了体外 XT 细胞系（CC/XT）。计划进行 CC/XT 的体内再接种（XT/CC）。通过形态学模式（苏木精/伊红）、免疫组织化学染色（CD99、FLI1、p16、p53、ki-67）、荧光原位杂交-FISH-（EWSR1 断裂分离、p16 和 p53 状态）和基因融合类型分析在裸鼠中生长的连续肿瘤传代的 TMA（XT/OT 和 XT/CC）。
OT、XT/OT、CC/XT 和 XT/CC 中的 p16、p53 和 ki67 观察到异质性结果。三个细胞系均显示 EWS/FLI1 重排。仅在一个病例中发现 p16 基因缺失。缺失通过 FISH 检测，并通过 PCR 检测证实。第二个病例在 CC 进化过程中发现 17 号染色体单体和随后的多体状态的 p53 改变。PCR 研究显示 p53 突变。第三个病例显示 p16 启动子的高甲基化。从 CC/XT 进行接种时，裸鼠中的肿瘤生长更迅速，随着传代的进行逐渐增加。在重新接种 CC/XT 后，第三个病例未显示裸鼠中的肿瘤生长。
在 TMA 中对原始肿瘤和连续肿瘤传代的多个核心进行研究，有助于分析 Ewing 肿瘤的遗传改变和蛋白质表达。
