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
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
复制标题
肿瘤坏死因子 α 诱导蛋白 1 作为一种新型肿瘤抑制因子，通过选择性下调 CSNK2B 阻断肝细胞癌中核因子-κ B 的激活
DOI：
10.1016/j.ebiom.2019.102603
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发表时间：
2020-01-01
期刊：
影响因子：
11.1
通讯作者：
Xiang，Shuanglin
中科院分区：
文献类型：
--
作者：
Xiao，Ye;Huang，Shulan;Xiang，Shuanglin
肿瘤坏死因子α诱导蛋白1在肿瘤细胞系中表达下调，并促进肿瘤细胞的凋亡。然而，其在肝细胞癌中的作用、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织化学方法检测TNFAIP1在肝癌组织和细胞系中的表达。采用细胞计数试剂盒(CCK8)、末端脱氧核苷酸转移酶dUTP缺口末端标记法(TUNEL)、Transwell法、裸鼠体内成管法等方法观察TNFAIP1对肝癌细胞增殖、凋亡、转移、血管生成和肿瘤形成的影响。用LC-MS/MS、免疫共沉淀和Western印迹等方法验证了TNFAIP1与CSNK2B之间的相互作用。采用双荧光素酶报告、免疫荧光、实时定量聚合酶链式反应(RT-κ)和免疫印迹等方法分析了TNFAIP1调控核转录因子-kappaB(NF-kappaB)途径的机制。在体内外，TNFAIP1的过表达抑制了肝癌细胞的增殖、转移、血管生成，促进了癌细胞的凋亡，而在肝癌细胞中过表达的TNFAIP1则表现出相反的作用。在机制上，TNFAIP1与CSNK2B相互作用，并促进其与CUL3的泛素介导的降解，导致肝癌细胞中依赖CSNK2B的NF-κB反式激活减弱。说明TNFAIP1可能通过调节TNFAIP1/CSNK2B/NF-κB信号通路发挥肿瘤抑制作用，提示TNFAIP1可能是一种潜在的肝癌标记物和治疗靶点。
BackgroundTumor 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.MethodsThe 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 assayin vitroand nude mice experimentsin 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.FindingsThe 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 bothin vitroandin 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 angiogenesisin vitroandin vivo.InterpretationOur 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.