AMD1 upregulates hepatocellular carcinoma cells stemness by FTO mediated mRNA demethylation.
AMD1 upregulates hepatocellular carcinoma cells stemness by FTO mediated mRNA demethylation.
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AMD1通过FTO介导mRNA去甲基化上调肝癌细胞干性
DOI：
10.1002/ctm2.352
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发表时间：
2021-03
影响因子：
10.6
通讯作者：
Wu W
中科院分区：
文献类型：
--
作者：
Bian X;Shi D;Xing K;Zhou H;Lu L;Yu D;Wu W
S‐腺苷蛋氨酸脱羧酶前酶（AMD1）是参与合成精胺（SPM）和亚精胺（SPD）的关键酶，与多种细胞过程相关。它也被发现是多种癌症的致癌基因和肿瘤治疗的潜在靶点。然而，AMD1在肝细胞癌（HCC）中的作用尚不清楚。应用肝细胞癌样本检测AMD1表达并评估其与临床病理特征和预后的关系。建立小鼠皮下和原位肿瘤模型，分析AMD1敲低或过表达后肝癌细胞的增殖和转移情况。采用药敏法和肿瘤球法观察AMD1对肝癌细胞干细胞性的影响。采用实时定量PCR （qRT‐PCR）、western blot、免疫组化（IHC）和m6A‐RNA免疫沉淀（Me‐RIP）测序/qPCR技术探讨AMD1在HCC中的潜在机制。此外，采用免疫荧光、co - IP （co - IP）和质谱（MS）分析来验证与AMD1相互作用的蛋白。AMD1在人HCC组织中富集，提示预后较差。高水平的AMD1可通过肥胖相关蛋白（FTO）介导的mRNA去甲基化促进SRY - box转录因子2 （SOX2）、Kruppel样因子4 （KLF4）和NANOG的表达。机制上，AMD1的高表达增加了HCC细胞中SPD的水平，从而修饰了支架蛋白Ras GTPase激活样蛋白1 (IQGAP1)，并增强了IQGAP1与FTO的相互作用。这种相互作用可以增强FTO的磷酸化，降低其泛素化。AMD1可以稳定IQGAP1与FTO的相互作用，从而促进FTO的表达，增加HCC的干细胞性。AMD1有望成为HCC的预后预测因子和治疗靶点。AMD1可以稳定IQGAP1与FTO的相互作用。与IQGAP1的相互作用增加了FTO的磷酸化和表达。高水平的FTO促进多能因子的表达，提高HCC细胞的干细胞样特性。
S‐adenosylmethionine decarboxylase proenzyme (AMD1) is a key enzyme involved in the synthesis of spermine (SPM) and spermidine (SPD), which are associated with multifarious cellular processes. It is also found to be an oncogene in multiple cancers and a potential target for tumor therapy. Nevertheless, the role AMD1 plays in hepatocellular carcinoma (HCC) is still unknown. HCC samples were applied to detect AMD1 expression and evaluate its associations with clinicopathological features and prognosis. Subcutaneous and orthotopic tumor mouse models were constructed to analyze the proliferation and metastasis of HCC cells after AMD1 knockdown or overexpression. Drug sensitive and tumor sphere assay were performed to investigate the effect of AMD1 on HCC cells stemness. Real‐time quantitative PCR (qRT‐PCR), western blot, immunohistochemical (IHC) and m6A‐RNA immunoprecipitation (Me‐RIP) sequencing/qPCR were applied to explore the potential mechanisms of AMD1 in HCC. Furthermore, immunofluorescence, co‐IP (Co‐IP) assays, and mass spectrometric (MS) analyses were performed to verify the proteins interacting with AMD1. AMD1 was enriched in human HCC tissues and suggested a poor prognosis. High AMD1 level could promote SRY‐box transcription factor 2 (SOX2), Kruppel like factor 4 (KLF4), and NANOG expression of HCC cells through obesity–associated protein (FTO)‐mediated mRNA demethylation. Mechanistically, high AMD1 expression increased the levels of SPD in HCC cells, which could modify the scaffold protein, Ras GTPase‐activating‐like protein 1 (IQGAP1) and enhance the interaction between IQGAP1 and FTO. This interaction could enhance the phosphorylation and decrease the ubiquitination of FTO. AMD1 could stabilize the interaction of IQGAP1 with FTO, which then promotes FTO expression and increases HCC stemness. AMD1 shows prospects as a prognostic predictor and a therapeutic target for HCC. AMD1 could stabilize the interaction of IQGAP1 with FTO. The interaction with IQGAP1 increases FTO phosphorylation and expression. High level of FTO promotes pluripotency factors expression and elevates stem cell‐like property of HCC cells.
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