MNAT1 is overexpressed in colorectal cancer and mediates p53 ubiquitin-degradation to promote colorectal cancer malignance.
MNAT1 is overexpressed in colorectal cancer and mediates p53 ubiquitin-degradation to promote colorectal cancer malignance.
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DOI：
10.1186/s13046-018-0956-3
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发表时间：
2018-11-26
期刊：
影响因子：
--
通讯作者：
Tang F
中科院分区：
文献类型：
--
作者：
Zhou S;Lu J;Li Y;Chen C;Cai Y;Tan G;Peng Z;Zhang Z;Dong Z;Kang T;Tang F
MNAT 1（menage a trois 1，MAT 1）是一种细胞周期蛋白依赖性激酶激活激酶（cyclin-dependent kinase-activating kinase，CAK）复合物，在多种肿瘤中高表达，参与肿瘤的发病机制。然而，其在致癌作用中的调节机制尚不清楚。应用大肠癌组织芯片，采用免疫组化方法检测大肠癌组织中MNAT 1的表达，并采用Western-blotting方法检测大肠癌细胞系中MNAT 1的表达。构建MNAT 1和shMNAT 1真核表达载体，转染大肠癌细胞。用MTT法和集落形成法观察转染细胞的生长情况。采用Western-blotting和Real-time PCR检测MNAT 1对p53表达的影响。免疫沉淀法检测p53与MNAT 1的相互作用，Western-blotting法检测MNAT 1对p53下游分子的影响。流式细胞仪检测MNAT 1和shMNAT 1对大肠癌细胞凋亡的影响。以BABL/c裸鼠为模型，观察MNAT 1对结直肠癌细胞生长的影响。发现MNAT 1在CRC组织和细胞中过表达，并且在CRC组织样本中的MNAT 1表达与CRC癌变和不良患者结局相关。MNAT 1敲入增加CRC细胞生长和集落形成，MNAT 1敲低显著降低细胞运动性和侵袭性。MNAT 1与p53物理相互作用，MNAT 1也增加MDM 2与p53的相互作用。引人注目的是，MNAT 1介导了p53遍在蛋白降解。MNAT 1缩短了p53的半衰期，并且异位MNAT 1表达降低了p53蛋白的稳定性。此外，MNAT 1诱导RAD 51并降低p21、切割的caspase 3、切割的PARP和BAX表达。MNAT 1抑制CRC细胞凋亡。在p53增加后，shMANT 1减少裸鼠肿瘤生长。MNAT 1与p53结合，通过MDM 2介导p53泛素降解，增加细胞生长，减少细胞凋亡，最终促进CRC的发生。MNAT 1与p53结合并介导p53泛素降解轴是p53通路中的一个新的分子关节。本文的在线版本（10.1186/s13046-018-0956-3）包含补充材料，可供授权用户使用。
MNAT1 (menage a trois 1, MAT1), a cyclin-dependent kinase-activating kinase (CAK) complex, high expresses in various cancers and is involved in cancer pathogenesis. However, mechanisms underlying its regulation in carcinogenesis are unclear. The tissue microarray of colorectal cancer (CRC) was used to evaluate MNAT1 expressions in CRC tissues using immunohistochemistry, CRC cell lines were also detected MNAT1 expression using Western-blotting. MNAT1 and shMNAT1 vectors were constructed, and transfected into CRC cells. Cell growths of the transfected cells were observed using MTT and colony formation. The affects of MNAT1 on p53 expression were analyzed using Western-blotting and Real-time PCR. Immunoprecipitation assay was used to analyze the interaction p53 and MNAT1, and Western-blotting was used to test the effects of MNAT1 on p53 downstream molecules. The apoptosis of CRC cells with MNAT1 or shMNAT1 were analyzed using flow cytometry. BABL/c athymic nude mice were used to observe the effect of MNAT1 on CRC cell growth in vivo. MNAT1 was found to be overexpressed in CRC tissues and cells, and MNAT1 expressions in CRC tissue samples were associated with CRC carcinogenesis and poor patient outcomes. MNAT1-knockin increased CRC cell growth and colony formation, and MNAT1-knockdown dramatically decreased cell motility and invasion. MNAT1 physically interacted with p53, MNAT1 also increased the interaction of MDM2 with p53. Strikingly, MNAT1 mediated p53 ubiquitin-degradation. MNAT1 shortened p53 half-life, and ectopic MNAT1 expression decreased p53 protein stability. Moreover, MNAT1 induced RAD51 and reduced p21, cleaved-caspase3, cleaved-PARP and BAX expression. MNAT1 inhibited CRC cell apoptosis. shMANT1 decreased tumor growths in nude mice following p53 increase. MNAT1 binds to p53, mediates p53 ubiquitin-degradation through MDM2, increases cell growth and decreases cell apoptosis, and finally promotes CRC malignance. MNAT1 binding to p53 and mediating p53 ubiquitin-degradation axis represents a novel molecular joint in the p53 pathway. The online version of this article (10.1186/s13046-018-0956-3) contains supplementary material, which is available to authorized users.
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