Lund Anders H, Turner Geoffrey, Trubetskoy Alla, Verhoeven Els, Wientjens Ellen, Hulsman Danielle, Russell Robert, DePinho Ronald A, Lenz Jack, van Lohuizen Maarten
Division of Molecular Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands.
Nat Genet. 2002 Sep;32(1):160-5. doi: 10.1038/ng956. Epub 2002 Aug 19.
We have used large-scale insertional mutagenesis to identify functional landmarks relevant to cancer in the recently completed mouse genome sequence. We infected Cdkn2a(-/-) mice with Moloney murine leukemia virus (MoMuLV) to screen for loci that can participate in tumorigenesis in collaboration with loss of the Cdkn2a-encoded tumor suppressors p16INK4a and p19ARF. Insertional mutagenesis by the latent retrovirus was synergistic with loss of Cdkn2a expression, as indicated by a marked acceleration in the development of both myeloid and lymphoid tumors. We isolated 747 unique sequences flanking retroviral integration sites and mapped them against the mouse genome sequence databases from Celera and Ensembl. In addition to 17 insertions targeting gene loci known to be cancer-related, we identified a total of 37 new common insertion sites (CISs), of which 8 encode components of signaling pathways that are involved in cancer. The effectiveness of large-scale insertional mutagenesis in a sensitized genetic background is demonstrated by the preference for activation of MAP kinase signaling, collaborating with Cdkn2a loss in generating the lymphoid and myeloid tumors. Collectively, our results show that large-scale retroviral insertional mutagenesis in genetically predisposed mice is useful both as a system for identifying genes underlying cancer and as a genetic framework for the assignment of such genes to specific oncogenic pathways.
我们利用大规模插入诱变技术，在最近完成的小鼠基因组序列中确定与癌症相关的功能标记。我们用莫洛尼氏鼠白血病病毒（MoMuLV）感染Cdkn2a(-/-)小鼠，以筛选那些能与Cdkn2a编码的肿瘤抑制因子p16INK4a和p19ARF缺失协同参与肿瘤发生的基因座。潜伏性逆转录病毒介导的插入诱变与Cdkn2a表达缺失具有协同作用，这在髓系和淋巴系肿瘤发展的显著加速中得到体现。我们分离出了747个位于逆转录病毒整合位点侧翼的独特序列，并将它们与来自Celera和Ensembl的小鼠基因组序列数据库进行比对。除了17个插入位点靶向已知与癌症相关的基因座外，我们总共鉴定出37个新的常见插入位点（CISs），其中8个编码参与癌症的信号通路成分。大规模插入诱变在敏感遗传背景下的有效性通过对MAP激酶信号激活的偏好得以证明，它与Cdkn2a缺失协同作用导致淋巴系和髓系肿瘤的发生。总体而言，我们的结果表明，在遗传易患性小鼠中进行大规模逆转录病毒插入诱变，既可用作鉴定癌症相关基因的系统，也可用作将此类基因分配到特定致癌途径的遗传框架。
