Wagner Kay-Uwe, Krempler Andrea, Qi Yongyue, Park KyungRan, Henry MaLinda D, Triplett Aleata A, Riedlinger Gregory, Rucker III Edmund B, Hennighausen Lothar
Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha 68198-6805, USA.
Mol Cell Biol. 2003 Jan;23(1):150-62. doi: 10.1128/MCB.23.1.150-162.2003.
Tumor susceptibility gene 101 (Tsg101) was identified in a random mutagenesis screen for potential tumor suppressors in NIH 3T3 cells. Altered transcripts of this gene have been detected in sporadic breast cancers and many other human malignancies. However, the involvement of this gene in neoplastic transformation and tumorigenesis is still elusive. Using gene targeting, we generated genetically engineered mice with a floxed allele of Tsg101. We investigated essential functions of this gene in vivo and examined whether the loss of function of Tsg101 results in tumorigenesis. Conventional knockout mice were generated through Cre-mediated excision of the first coding exon in the germ line of mouse mammary tumor virus (MMTV)-Cre transgenic mice. The complete ablation of Tsg101 in the developing embryo resulted in death around implantation. In contrast, mammary gland-specific knockout mice developed normally but were unable to nurse their young as a result of impaired mammogenesis during late pregnancy. Neither heterozygous null mutants nor somatic knockout mice developed mammary tumors after a latency of 2 years. The Cre-mediated deletion of Tsg101 in primary cells demonstrated that this gene is essential for the growth, proliferation, and survival of mammary epithelial cells. In summary, our results suggest that Tsg101 is required for normal cell function of embryonic and adult tissues but that this gene is not a tumor suppressor for sporadic forms of breast cancer.
肿瘤易感基因101(Tsg101)是在对NIH 3T3细胞中潜在肿瘤抑制因子的随机诱变筛选中被鉴定出来的。在散发性乳腺癌和许多其他人类恶性肿瘤中已检测到该基因的转录本改变。然而,该基因在肿瘤转化和肿瘤发生中的作用仍不明确。我们利用基因打靶技术,构建了Tsg101等位基因带有loxP位点的基因工程小鼠。我们研究了该基因在体内的基本功能,并检测Tsg101功能缺失是否会导致肿瘤发生。通过在小鼠乳腺肿瘤病毒(MMTV)-Cre转基因小鼠的种系中经Cre介导切除第一个编码外显子,产生了传统的基因敲除小鼠。发育中的胚胎中Tsg101的完全缺失导致在着床前后死亡。相比之下,乳腺特异性敲除小鼠发育正常,但由于妊娠后期乳腺发育受损而无法哺育幼崽。杂合性无效突变体和体细胞敲除小鼠在2年的潜伏期后均未发生乳腺肿瘤。在原代细胞中经Cre介导删除Tsg101表明,该基因对乳腺上皮细胞的生长、增殖和存活至关重要。总之,我们的结果表明,Tsg101是胚胎和成年组织正常细胞功能所必需的,但该基因不是散发性乳腺癌的肿瘤抑制因子。