Laboratory of Molecular Pathology, Department of Pathology, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy.
Genes Chromosomes Cancer. 2012 Feb;51(2):111-26. doi: 10.1002/gcc.20933. Epub 2011 Nov 1.
To highlight possible similarities and differences in receptor tyrosine kinase (RTK) and downstream signalling activation profiles between clear-cell sarcomas (CCS) and metastatic melanomas (MM), frozen, and paired-matched fixed samples of six CCS with EWSR1 rearrangement (EWSR1+), five CCS without EWSR1 rearrangement (EWSR1-), and seven MM were investigated by means of biochemical, immunohistochemical, FISH, molecular analyses, and immunofluorescence confocal microscopy. Fixed samples of a further 10 CCS and 14 MM were investigated by means of sequencing for BRAF, NRAS, and KRAS mutations and FISH analyses for the gain of chromosomes 22 and 8. RTK analysis of all CCS/MM samples showed activation of short-form (sf) recepteur d'origine nantais (RON) RTK and of PDGFRB, MET, and HER3. Analysis of downstream signaling revealed consistent phosphorylation patterns of PI3K/AKT, RSK, and the mTOR targets S6 and 4EBP1. Analysis of frozen and fixed material from 21 CCS and 21 MM showed the presence of the V600E BRAF mutation in 2/12 EWSR1+ and 3/9 EWSR1- CCS and 9/21 MM and demonstrated a significant (P < 0.001) correlation between the gain of chromosomes 22 and 8 and EWSR1- CCS. Our results show that BRAF mutation can also be present in CCS and support the proposed aberration of chromosomes 22 and 8 as a possibly useful nonrandom hallmark of EWSR1- CCS. Besides, they broaden the spectrum of the similarities of RTK pathway activation between CCS and MM, thus suggesting that new drugs found to be active in melanoma and RON inhibitors could have a role in CCS treatment. © 2011 Wiley Periodicals, Inc.
为了突出 EWSR1 重排的透明细胞肉瘤 (CCS) 和转移性黑色素瘤 (MM) 之间受体酪氨酸激酶 (RTK) 和下游信号激活谱的可能相似性和差异，我们通过生物化学、免疫组织化学、FISH、分子分析和免疫荧光共聚焦显微镜，对 6 例 EWSR1+CCS、5 例 EWSR1-CCS 和 7 例 MM 的冷冻和配对固定样本进行了研究。我们通过测序 BRAF、NRAS 和 KRAS 突变和 FISH 分析 22 号和 8 号染色体获得，对另外 10 例 CCS 和 14 例 MM 的固定样本进行了研究。所有 CCS/MM 样本的 RTK 分析显示短形式 (sf) recepteur d'origine nantais (RON) RTK 和 PDGFRB、MET 和 HER3 的激活。下游信号分析显示 PI3K/AKT、RSK 和 mTOR 靶标 S6 和 4EBP1 的一致磷酸化模式。对 21 例 CCS 和 21 例 MM 的冷冻和固定材料的分析显示，在 12 例 EWSR1+和 9 例 EWSR1-CCS 中的 2/12 例和 21 例 MM 中的 9/21 例中存在 V600E BRAF 突变，并证明 22 号和 8 号染色体的获得与 EWSR1-CCS 之间存在显著的相关性（P <0.001）。我们的结果表明，BRAF 突变也可能存在于 CCS 中，并支持染色体 22 和 8 的异常作为 EWSR1-CCS 的一个可能有用的非随机标志。此外，它们拓宽了 CCS 和 MM 之间 RTK 通路激活的相似性范围，从而表明在黑色素瘤中发现的新药物和 RON 抑制剂可能在 CCS 治疗中发挥作用。
