ANKHD1 is required for SMYD3 to promote tumor metastasis in hepatocellular carcinoma
ANKHD1 is required for SMYD3 to promote tumor metastasis in hepatocellular carcinoma
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ANKHD1是SMYD3促进肝细胞癌肿瘤转移所必需的
DOI：
10.1186/s13046-018-1011-0
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发表时间：
2019
影响因子：
11.3
通讯作者：
He Chuanchao
中科院分区：
文献类型：
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作者：
Zhou Zhenyu;Jiang Hai;Tu Kangsheng;Yu Wei;Zhang Jianlong;Hu Zhigang;Zhang Heyun;Hao Dake;Huang Pinbo;Wang Jie;Wang Aijun;Xiao Zhiyu;He Chuanchao
肿瘤转移是肝细胞癌患者肝切除术后预后不良的主要原因。SMYD3已被证明能促进小鼠肝肿瘤的转移。方法采用免疫组织化学、迁移实验、侵袭实验、伤口愈合实验和体内肺转移实验等方法分析SMYD3对肝癌细胞侵袭转移的影响。用H3K4me3抗体在SMYD3过表达细胞中拉下的蛋白质进行质谱分析。用荧光素酶报告基因、染色质免疫沉淀、凝胶迁移率改变实验检测SMYD3-ANKHD1对SLUG转录的调控。此外，采用免疫组织化学方法检测了243例肝癌组织中SMYD3-ANKHD1的表达情况。结果SMYD3是一种独立的肝癌预后因子，可促进肝癌细胞的迁移和侵袭。质谱学鉴定ANKHD1与SMYD3共同调控。当细胞过度表达SMYD3时，ANKHD1与H3K4me3相互作用。当ANKHD1被siRNA击倒时，SMYD3的亲迁移和亲侵袭作用被减弱。此外，我们还发现SMYD3结合和激活SLUG基因启动子的方式与H3K4me3、H3K9Ac和H3K14Ac的升高有关。敲除ANKHD1可以减弱SMYD3依赖的Slug表达的激活。我们进一步检测了243例肝癌组织中SMYD3和ANKHD1的表达，发现SMYD3和ANKHD1共表达阳性的患者总体生存时间最短，无复发生存期最短。结论SMYD3-ANKHD1基因为SMYD3调控肝癌转移提供了一种新的分子机制，提示SMYD3-ANKHD1可能成为治疗肝癌的潜在靶点。
BackgroundTumor metastasis is the major reason for poor prognosis of hepatocellular carcinoma (HCC) patients after hepatic resection. SMYD3 has been demonstrated to promote liver tumor metastasis in mice. However, the detailed molecular mechanism is still largely unknown.MethodsThe effect of SMYD3 on invasiveness and metastasis of HCC was analyzed by immunohistochemistry, migration assay, invasion assay, wound healing assay and in vivo lung metastasis assay. Mass spectrometry analysis was conducted using proteins pulled down by H3K4me3 antibody in SMYD3-overexpressing cells. Luciferase reporter, chromatin immunoprecipitation, Electrophoretic mobility shift assay were used to measure the regulation ofSLUGtranscription by SMYD3-ANKHD1. In addition, the role of SMYD3-ANKHD1 in determining clinical outcomes for HCC patients was investigated by immunohistochemistry in 243 HCC tissues.ResultsSMYD3 was an independent prognostic factor of HCC and promoted migration and invasion of human HCC cells. ANKHD1 was identified by mass spectrometry as a co-regulator with SMYD3. ANKHD1 interacted with H3K4me3 when cells were overexpressing SMYD3. The pro-migratory and pro-invasive effects of SMYD3 were attenuated when ANKHD1 was knocked down by siRNA. Furthermore, we found that SMYD3 bound and activated theSLUGgene promoter in a manner associated with elevating H3K4me3, H3K9Ac and H3K14Ac. Knockdown of ANKHD1 could attenuate the SMYD3-dependent activation of Slug expression. We further detected the expression of SMYD3 and ANKHD1 in 243 HCC patients and found that patients with positive coexpression of SMYD3 and ANKHD1 (SMYD3+ANKHD1+) had the shortest overall and recurrence-free survival.ConclusionOur findings provide a novel molecular mechanism for the SMYD3-regulated HCC migration and metastasis, and indicates that SMYD3-ANKHD1 may be a potential target for treating HCC.