A novel Notch1 missense mutation (C1133Y) in the Abruptex domain exhibits enhanced proliferation and invasion in oral squamous cell carcinoma.
A novel Notch1 missense mutation (C1133Y) in the Abruptex domain exhibits enhanced proliferation and invasion in oral squamous cell carcinoma.
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Abruptex 结构域中的新型 Notch1 错义突变 (C1133Y) 表现出口腔鳞状细胞癌的增殖和侵袭增强
DOI：
10.1186/s12935-017-0496-5
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发表时间：
2018
影响因子：
5.8
通讯作者：
Song X
中科院分区：
文献类型：
--
作者：
Zheng Y;Wang Z;Ding X;Zhang W;Li G;Liu L;Wu H;Gu W;Wu Y;Song X
背景Notch 1被认为是组织分化和干细胞特性的基本调节因子。最近，Notch 1突变在实体瘤和造血系统恶性肿瘤中都有大量报道。然而，很少有人知道这些报告的突变的生物学效应和临床意义。以前，我们发现了几个错义突变的Notch 1受体在中国人口与口腔鳞状细胞癌（OSCC）。MethodsWe选择了一个“热点”突变的Abruptex域（C1133 Y）。Western blot和实时定量PCR检测转染pcDNA3.1-Notch 1 WT、pcDNA3.1-Notch 1C 1133 Y和pcDNA3.1空载体的OSCC细胞中Notch 1的表达。CCK-8测定用于评估细胞增殖。流式细胞术和western blot检测转染后细胞周期的变化。采用Transwell法和上皮间质转化（EMT）标志物检测细胞的侵袭能力。通过免疫荧光染色和EGFR-PI 3 K/AKT信号通路的表达分析Notch 1C 1133 Y突变的影响。我们通过促进细胞增殖、侵袭和诱导OSCC细胞系中的EMT来鉴定这种突变的致癌表型。我们发现，Notch 1C 1133 Y突变表现出减少的S1-切割由于受损的运输Notch 1蛋白从内质网（ER）的高尔基复合体，这是一致的观察失败的Notch 1C 1133 Y突变的受体目前在细胞表面。重要的是，突变的Notch 1激活EGFR-PI 3 K/AKT信号通路，这已被证实为一个压倒性的调制器在OSCC.ConclusionsTaken在一起，我们的研究结果首次揭示了一种新的Notch 1突变，增强增殖和侵袭的OSCC细胞系。Notch 1 C1133 Y突变损害了Notch 1蛋白的加工以及突变的Notch 1与活化的EGFR-PI 3 K/AKT信号通路之间的关键联系。
BackgroundNotch1 has been regarded as a fundamental regulator in tissue differentiation and stem cell properties. Recently, Notch1 mutations have been reported intensively both in solid tumors and in hematopoietic malignancies. However, little is known about the biological effect and the clinical implication of these reported mutations. Previously, we discovered several missense mutations in the Notch1 receptor in a Chinese population with oral squamous cell carcinoma (OSCC).MethodsWe selected a ‘hotspot’ mutation in the Abruptex domain (C1133Y). The expression of Notch1 was determined by western blot and real-time qPCR in OSCC cell lines transfected with pcDNA3.1-Notch1WT, pcDNA3.1-Notch1C1133Y, or pcDNA3.1 empty vector. CCK-8 assays were used to assess cell proliferation. Flow cytometry and western blot were used to confirm the alteration of cell cycle after transfection. Transwell assays and the detection of Epithelial-to-mesenchymal transition (EMT) markers were used to determine the invasive ability. The effects of Notch1 C1133Y mutation were analyzed by Immunofluorescence staining and the expression of EGFR-PI3K/AKT signaling.ResultsWe demonstrated that Notch1C1133Ymutation inactivated the canonical Notch1 signaling. We identified an oncogenic phenotype of this mutation by promoting cell proliferation, invasion and by inducing EMT in OSCC cell lines. We found that the Notch1C1133Ymutation exhibited a decreased S1-cleavage due to the impaired transport of Notch1 protein from the endoplasmic reticulum (ER) to the Golgi complex, which was consistent with the observation of the failure of the Notch1C1133Ymutated receptor to present at the cell surface. Importantly, the mutated Notch1 activated the EGFR-PI3K/AKT signaling pathway, which has been confirmed as an overwhelming modulator in OSCC.ConclusionsTaken together, our findings revealed for the first time a novel Notch1 mutation that enhances proliferation and invasion in OSCC cell lines. The Notch1 C1133Y mutation impairs the processing of notch1 protein and the critical links between the mutated Notch1 and the activated EGFR-PI3K/AKT signaling pathway.
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影响因子：
7.3
作者：
Gonzalez DM;Medici D
通讯作者：
Medici D
影响因子：
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作者：
Jimenez L;Jayakar SK;Ow TJ;Segall JE
通讯作者：
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作者：
Dai, Jianjian;Ma, Daoxin;Ji, Chunyan
通讯作者：
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影响因子：
64.5
作者：
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通讯作者：
YOUNG, MW
影响因子：
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作者：
Chang, Alex C. Y.;Garside, Victoria C.;Karsan, Aly
通讯作者：
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