Department of Immunology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
1] Department of Immunology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA [2] The University of Texas Graduate School of Biomedical Sciences at Houston, Houston, TX, USA.
Oncogene. 2014 May 22;33(21):2748-57. doi: 10.1038/onc.2013.234. Epub 2013 Jun 10.
Ku70, a known nonhomologous end-joining (NHEJ) factor, also functions in tumor suppression, although this molecular mechanism remains uncharacterized. Previously, we showed that mice deficient for DNA ligase IV (Lig4), another key NHEJ factor, succumbed to aggressive lymphoma in the absence of tumor suppressor p53. However, the tumor phenotype is abrogated by the introduction of a hypomorphic mutant p53(R172P), which impaired p53-mediated apoptosis but not cell-cycle arrest. However, Lig4(-/-)p53(R172P) mice succumbed to severe diabetes. To further elucidate the role of NHEJ and p53-mediated apoptosis in vivo, we bred Ku70(-/-) p53(R172P) mice. Unexpectedly, these mice were free of diabetes, although 80% of the mutant mice had abnormally enlarged colons with pronounced inflammation. Remarkably, most of these mutant mice progressed to dysplasia, adenoma and adenocarcinoma; this is in contrast to the Lig4(-/-)p53(R172P) phenotype, strongly suggesting an NHEJ-independent function of Ku70. Significantly, our analyses of Ku70(-/-)p53(R172P) colonic epithelial cells show nuclear stabilization of β-catenin accompanied by higher expression of cyclin D1 and c-Myc in affected colon sections than in control samples. This is not due to the p53 mutation, as Ku70(-/-) mice share this phenotype. Our results not only unravel a novel function of Ku70 essential for colon homeostasis, but also establish an excellent in vivo model in which to study how chronic inflammation and abnormal cellular proliferation underlie tumorigenesis and tumor progression in the colon.
Ku70 是一种已知的非同源末端连接(NHEJ)因子,也具有肿瘤抑制功能,尽管其分子机制尚不清楚。以前,我们曾表明,另一种关键 NHEJ 因子 DNA 连接酶 IV(Lig4)缺失的小鼠在没有肿瘤抑制因子 p53 的情况下会患上侵袭性淋巴瘤。然而,引入一个功能减弱的 p53(R172P)突变体可消除肿瘤表型,该突变体削弱了 p53 介导的细胞凋亡,但不影响细胞周期阻滞。然而,Lig4(-/-)p53(R172P)小鼠却患上了严重的糖尿病。为了进一步阐明 NHEJ 和 p53 介导的细胞凋亡在体内的作用,我们培育了 Ku70(-/-) p53(R172P)小鼠。出乎意料的是,这些小鼠没有糖尿病,尽管 80%的突变小鼠的结肠异常增大,伴有明显的炎症。值得注意的是,大多数这些突变小鼠进展为异型增生、腺瘤和腺癌;这与 Lig4(-/-)p53(R172P)的表型形成鲜明对比,强烈表明 Ku70 具有非依赖 NHEJ 的功能。重要的是,我们对 Ku70(-/-) p53(R172P)结肠上皮细胞的分析表明,β-catenin 在细胞核中的稳定性增加,同时受影响的结肠切片中 cyclin D1 和 c-Myc 的表达高于对照样本。这不是由于 p53 突变所致,因为 Ku70(-/-)小鼠具有这种表型。我们的研究结果不仅揭示了 Ku70 对于结肠稳态所必需的一种新功能,而且建立了一个极好的体内模型,可用于研究慢性炎症和异常细胞增殖如何导致结肠肿瘤发生和肿瘤进展。