Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta, Canada.
Am J Pathol. 2011 Jul;179(1):411-21. doi: 10.1016/j.ajpath.2011.03.045. Epub 2011 May 24.
The fusion tyrosine kinase NPM-ALK is central to the pathogenesis of ALK-positive anaplastic large cell lymphoma (ALK(+)ALCL). We recently identified that MSH2, a key DNA mismatch repair (MMR) protein integral to the suppression of tumorigenesis, is an NPM-ALK-interacting protein. In this study, we found in vitro evidence that enforced expression of NPM-ALK in HEK293 cells suppressed MMR function. Correlating with these findings, six of nine ALK(+)ALCL tumors displayed evidence of microsatellite instability, as opposed to none of the eight normal DNA control samples (P = 0.007, Student's t-test). Using co-immunoprecipitation, we found that increasing levels of NPM-ALK expression in HEK293 cells resulted in decreased levels of MSH6 bound to MSH2, whereas MSH2·NPM-ALK binding was increased. The NPM-ALK·MSH2 interaction was dependent on the activation/autophosphorylation of NPM-ALK, and the Y191 residue of NPM-ALK was a crucial site for this interaction and NPM-ALK-mediated MMR suppression. MSH2 was found to be tyrosine phosphorylated in the presence of NPM-ALK. Finally, NPM-ALK impeded the expected DNA damage-induced translocation of MSH2 out of the cytoplasm. To conclude, our data support a model in which the suppression of MMR by NPM-ALK is attributed to its ability to interfere with normal MSH2 biochemistry and function.
核仁磷酸蛋白 NPM-ALK 是间变性大细胞淋巴瘤(ALK(+)ALCL)发病机制的核心。我们最近发现,错配修复(MMR)关键蛋白 MSH2 是一种抑制肿瘤发生的必需蛋白,它是 NPM-ALK 的相互作用蛋白。在这项研究中,我们在体外发现,NPM-ALK 在 HEK293 细胞中的强制表达抑制了 MMR 功能。与这些发现相关的是,9 例 ALK(+)ALCL 肿瘤中有 6 例显示微卫星不稳定的证据,而 8 例正常 DNA 对照样本中无一例显示(P = 0.007,Student's t 检验)。通过共免疫沉淀,我们发现,随着 HEK293 细胞中 NPM-ALK 表达水平的增加,与 MSH2 结合的 MSH6 水平下降,而 MSH2·NPM-ALK 结合增加。NPM-ALK·MSH2 相互作用依赖于 NPM-ALK 的激活/自磷酸化,并且 NPM-ALK 的 Y191 残基是这种相互作用和 NPM-ALK 介导的 MMR 抑制的关键位点。在存在 NPM-ALK 的情况下,MSH2 被酪氨酸磷酸化。最后,NPM-ALK 阻碍了 MSH2 预期的因 DNA 损伤而从细胞质中转位。总之,我们的数据支持这样一种模型,即 NPM-ALK 对 MMR 的抑制归因于其干扰正常 MSH2 生化和功能的能力。