A novel chemical screening strategy in zebrafish identifies common pathways in embryogenesis and rhabdomyosarcoma development
A novel chemical screening strategy in zebrafish identifies common pathways in embryogenesis and rhabdomyosarcoma development
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DOI：
10.1242/dev.088427
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发表时间：
2013-06-01
期刊：
影响因子：
4.6
通讯作者：
Zon, Leonard I.
中科院分区：
文献类型：
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作者：
Le, Xiuning;Pugach, Emily K.;Zon, Leonard I.
斑马鱼是一个强大的遗传模型，最近才被用来解剖肿瘤发生中涉及的发育途径。我们假设胚胎发生过程中的操作途径也会用于肿瘤发生。为了在胚胎发生期间定义RAS靶基因，在经受热休克的Tg（hsp 70-HRAS（G12 V））斑马鱼胚胎中评估基因表达。dusp 6被RAS激活，这被用作化学遗传筛选的基础，以鉴定在胚胎发生期间干扰RAS信号传导的小分子。然后使用KRAS（G12 D）诱导的斑马鱼胚胎横纹肌肉瘤来评估小分子的治疗效果。这些抑制剂中的两种，PD 98059和TPCK，在斑马鱼胚胎横纹肌肉瘤和NRAS中含有激活突变的横纹肌肉瘤的人细胞系中作为单一药剂具有抗肿瘤活性。PD 98059抑制MEK 1，而TPCK抑制S6 K1活性;然而，联合治疗完全抑制eIF 4 B磷酸化并降低翻译起始。我们的工作表明，在胚胎发生过程中RAS诱导的激活途径在肿瘤发生中也很重要，抑制这些途径可抑制肿瘤生长。
The zebrafish is a powerful genetic model that has only recently been used to dissect developmental pathways involved in oncogenesis. We hypothesized that operative pathways during embryogenesis would also be used for oncogenesis. In an effort to define RAS target genes during embryogenesis, gene expression was evaluated in Tg(hsp70-HRAS(G12V)) zebrafish embryos subjected to heat shock. dusp6 was activated by RAS, and this was used as the basis for a chemical genetic screen to identify small molecules that interfere with RAS signaling during embryogenesis. A KRAS(G12D)-induced zebrafish embryonal rhabdomyosarcoma was then used to assess the therapeutic effects of the small molecules. Two of these inhibitors, PD98059 and TPCK, had anti-tumor activity as single agents in both zebrafish embryonal rhabdomyosarcoma and a human cell line of rhabdomyosarcoma that harbored activated mutations in NRAS. PD98059 inhibited MEK1 whereas TPCK suppressed S6K1 activity; however, the combined treatment completely suppressed eIF4B phosphorylation and decreased translation initiation. Our work demonstrates that the activated pathways in RAS induction during embryogenesis are also important in oncogenesis and that inhibition of these pathways suppresses tumor growth.