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Tumor necrosis factor α-induced protein 1 as a novel tumor suppressor through selective downregulation of CSNK2B blocks nuclear factor-κB activation in hepatocellular carcinoma.
EBioMedicine ( IF 11.1 ) Pub Date : 2020-01-02 , DOI: 10.1016/j.ebiom.2019.102603
Ye Xiao 1 , Shulan Huang 2 , Feng Qiu 2 , Xiaofeng Ding 3 , Yi Sun 4 , Chenxi Wei 3 , Xiang Hu 3 , Ke Wei 5 , Shengwen Long 2 , Lina Xie 6 , Yu Xun 2 , Wen Chen 2 , Zhijian Zhang 7 , Ning Liu 8 , Shuanglin Xiang 3
Affiliation  

BACKGROUND Tumor necrosis factor α-induced protein 1 (TNFAIP1) is frequently downregulated in cancer cell lines and promotes cancer cell apoptosis. However, its role, clinical significance and molecular mechanisms in hepatocellular carcinoma (HCC) are unknown. METHODS The expression of TNFAIP1 in HCC tumor tissues and cell lines was measured by Western blot and immunohistochemistry. The effects of TNFAIP1 on HCC proliferation, apoptosis, metastasis, angiogenesis and tumor formation were evaluated by Cell Counting Kit-8 (CCK8), Terminal deoxynucleotidyl transferase dUTP Nick-End Labeling (TUNEL), transwell, tube formation assay in vitro and nude mice experiments in vivo. The interaction between TNFAIP1 and CSNK2B was validated by liquid chromatography-tandem mass spectrometry (LC-MS/MS), Co-immunoprecipitation and Western blot. The mechanism of how TNFAIP1 regulated nuclear factor-kappaB (NF-κB) pathway was analyzed by dual-luciferase reporter, immunofluorescence, quantitative Real-time polymerase chain reaction (RT-qPCR) and Western blot. FINDINGS The TNFAIP1 expression is significantly decreased in HCC tissues and cell lines, and negatively correlated with the increased HCC histological grade. Overexpression of TNFAIP1 inhibits HCC cell proliferation, metastasis, angiogenesis and promotes cancer cell apoptosis both in vitro and in vivo, whereas the knockdown of TNFAIP1 in HCC cell displays opposite effects. Mechanistically, TNFAIP1 interacts with CSNK2B and promotes its ubiquitin-mediated degradation with Cul3, causing attenuation of CSNK2B-dependent NF-κB trans-activation in HCC cell. Moreover, the enforced expression of CSNK2B counteracts the inhibitory effects of TNFAIP1 on HCC cell proliferation, migration, and angiogenesis in vitro and in vivo. INTERPRETATION Our results support that TNFAIP1 can act as a tumor suppressor of HCC by modulating TNFAIP1/CSNK2B/NF-κB pathway, implying that TNFAIP1 may represent a potential marker and a promising therapeutic target for HCC.

中文翻译:

肿瘤坏死因子α诱导蛋白1通过选择性下调CSNK2B来抑制肝细胞癌中核因子-κB的活化,是一种新型的肿瘤抑制因子。

背景技术肿瘤坏死因子α诱导的蛋白1(TNFAIP1)在癌细胞系中经常被下调并促进癌细胞凋亡。但是,其在肝细胞癌(HCC)中的作用,临床意义和分子机制尚不清楚。方法采用免疫印迹和免疫组化方法检测肝癌组织和细胞株中TNFAIP1的表达。通过细胞计数试剂盒8(CCK8),末端脱氧核苷酸转移酶dUTP尼克末端标记(TUNEL),transwell,体外管形成试验和裸鼠来评估TNFAIP1对HCC增殖,凋亡,转移,血管生成和肿瘤形成的影响体内实验。TNFAIP1和CSNK2B之间的相互作用已通过液相色谱-串联质谱(LC-MS / MS),共免疫沉淀和Western blot进行了验证。通过双荧光素酶报告基因,免疫荧光,实时定量聚合酶链反应(RT-qPCR)和蛋白质印迹分析了TNFAIP1调节核因子-κB(NF-κB)途径的机制。结果在肝癌组织和细胞系中,TNFAIP1的表达显着降低,并且与肝癌组织学分级的升高呈负相关。TNFAIP1的过表达在体外和体内均抑制HCC细胞的增殖,转移,血管生成并促进癌细胞凋亡,而在HCC细胞中敲低TNFAIP1则表现出相反的作用。从机制上讲,TNFAIP1与CSNK2B相互作用并促进其泛素介导的Cul3降解,从而导致HCC细胞中CSNK2B依赖性NF-κB反式激活的减弱。此外,CSNK2B的强制表达抵消了TNFAIP1在体外和体内对HCC细胞增殖,迁移和血管生成的抑制作用。解释我们的结果支持TNFAIP1可以通过调节TNFAIP1 / CSNK2B /NF-κB途径来作为HCC的肿瘤抑制因子,这暗示TNFAIP1可能代表了HCC的潜在标志物和有希望的治疗靶点。
更新日期:2020-01-04
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