FOXC1 promotes HCC proliferation and metastasis by Upregulating DNMT3B to induce DNA Hypermethylation of CTH promoter.
FOXC1 promotes HCC proliferation and metastasis by Upregulating DNMT3B to induce DNA Hypermethylation of CTH promoter.
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FOXC1 通过上调 DNMT3B 诱导 CTH 启动子 DNA 高甲基化来促进 HCC 增殖和转移。
DOI：
10.1186/s13046-021-01829-6
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发表时间：
2021-02-01
期刊：
影响因子：
--
通讯作者：
Xia L
中科院分区：
文献类型：
--
作者：
Lin Z;Huang W;He Q;Li D;Wang Z;Feng Y;Liu D;Zhang T;Wang Y;Xie M;Ji X;Sun M;Tian D;Xia L
叉头盒C1（FOXC 1）作为FOX家族的一员，在促进肝癌侵袭和转移中起重要作用。FOX家族蛋白在新陈代谢中起着关键作用。ROS参与肿瘤的发生发展，并与多种转录因子的表达相关。我们接下来探索了FOXC 1调节HCC中代谢和ROS止血的潜在机制。我们使用氨基酸阵列来验证哪些代谢参与FOXC 1诱导的HCC。用该试剂盒检测FOXC 1过表达或低表达的HCC细胞中ROS水平。在体内外筛选出FOXC 1调控肝癌进展的下游靶基因和候选通路后，采用免疫印迹、免疫组化、亚硫酸氢盐基因组测序、甲基化特异性PCR、染色质免疫沉淀分析和荧光素酶报告基因分析等方法，探讨FOXC 1与下游基因的关系。此外，在两个独立的人类HCC队列中分析FOXC 1与靶基因之间的相关性以及靶基因与复发和总生存率之间的相关性。本文报道了FOXC 1可通过调节半胱氨酸代谢相关基因胱硫醚γ-裂解酶（cystathioprotein γ-lyase，CTH）来抑制半胱氨酸代谢，增加活性氧（reactive oxygen species，ROS）水平。CTH过表达可显著抑制FOXC 1诱导的肝癌细胞增殖、侵袭和转移，而抑制FOXC 1所导致的细胞增殖、侵袭和转移的降低可被CTH敲低所逆转。同时，FOXC 1上调DNA甲基化酶3B（DNMT3B）的表达，诱导CTH基因启动子甲基化，导致CTH基因在肝癌细胞中的低表达。此外，由N-乙酰半胱氨酸（NAC），这是一种抗氧化剂诱导的低水平的ROS抑制FOXC 1过表达介导的细胞增殖，迁移和侵袭能力，而由L-丁硫氨酸-亚砜亚胺（BSO）诱导的高水平的ROS拯救FOXC 1敲低介导的抑制结果。我们的研究表明，由ROS诱导的FOXC 1的过表达依赖于细胞外调节蛋白激酶1和2（ERK 1/2）-磷酸化ETS转录因子1（p-ELK 1）途径。在人肝癌组织中，FOXC 1的表达与氧化损伤标志物8-羟基脱氧鸟苷（8-OHdG）、p-ELK 1和DNMT 3B的表达呈正相关，与CTH的表达呈负相关。8-OHdG/FOXC 1、p-ELK 1/FOXC 1、FOXC 1/DNMT 3B共表达的HCC患者预后最差，而FOXC 1阳性而CTH阴性的HCC患者预后最差。综上所述，我们阐明了ROS-FOXC 1-半胱氨酸代谢-ROS的正反馈环在促进肝癌细胞增殖和转移中的重要作用，该通路可能为肝癌的临床治疗提供一条有前景的途径。在线版本包含补充材料，可通过10.1186/s13046 - 021 - 01829 - 6获得。
Forkhead box C1 (FOXC1), as a member of the FOX family, is important for promote HCC invasion and metastasis. FOX family protein lays a pivotal role in metabolism. ROS is involved in tumor progression and is associated with the expression of lots of transcription factors. We next explored the mechanism underlying FOXC1 modulating the metabolism and ROS hemostasis in HCC. We used amino acids arrays to verify which metabolism is involved in FOXC1-induced HCC. The kits were used to detect the ROS levels in HCC cells with over-expression or down-expression of FOXC1. After identified the downstream target genes and candidate pathway which regulated by FOXC1 during HCC progression in vitro and in vivo, we used western blot, immunohistochemistry, bisulfite genomic sequencing, methylation-specific PCR, chromatin immunoprecipitation analysis and luciferase reporter assays to explore the relationship of FOXC1 and downstream genes. Moreover, the correlation between FOXC1 and target genes and the correlation between target genes and the recurrence and overall survival were analyzed in two independent human HCC cohorts. Here, we reported that FOXC1 could inhibit the cysteine metabolism and increase reactive oxygen species (ROS) levels by regulating cysteine metabolism-related genes, cystathionine γ-lyase (CTH). Overexpression of CTH significantly suppressed FOXC1-induced HCC proliferation, invasion and metastasis, while the reduction in cell proliferation, invasion and metastasis caused by the inhibition of FOXC1 could be reversed by knockdown of CTH. Meanwhile, FOXC1 upregulated de novo DNA methylase 3B (DNMT3B) expression to induce DNA hypermethylation of CTH promoter, which resulted in low expression of CTH in HCC cells. Moreover, low levels of ROS induced by N-acetylcysteine (NAC) which is an antioxidant inhibited the cell proliferation, migration, and invasion abilities mediated by FOXC1 overexpression, whereas high levels of ROS induced by L-Buthionine-sulfoximine (BSO) rescued the suppression results mediated by FOXC1 knockdown. Our study demonstrated that the overexpression of FOXC1 that was induced by the ROS dependent on the extracellular regulated protein kinases 1 and 2 (ERK1/2)- phospho-ETS Transcription Factor 1 (p-ELK1) pathway. In human HCC tissues, FOXC1 expression was positively correlated with oxidative damage marker 8-hydroxy-2′-deoxyguanosine (8-OHdG), p-ELK1 and DNMT3B expression, but negatively correlated with CTH expression. HCC patients with positive co-expression of 8-OHdG/FOXC1 or p-ELK1/FOXC1 or FOXC1/DNMT3B had the worst prognosis, whereas HCC patients who had positive FOXC1 and negative CTH expression exhibited the worst prognosis. In a word, we clarify that the positive feedback loop of ROS-FOXC1-cysteine metabolism-ROS is important for promoting liver cancer proliferation and metastasis, and this pathway may provide a prospective clinical treatment approach for HCC. The online version contains supplementary material available at 10.1186/s13046-021-01829-6.
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影响因子：
--
作者：
Kolukula VK;Sahu G;Wellstein A;Rodriguez OC;Preet A;Iacobazzi V;D'Orazi G;Albanese C;Palmieri F;Avantaggiati ML
通讯作者：
Avantaggiati ML
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DOI：
10.1186/s13046-017-0646-6
发表时间：
2017-12-06
期刊：
Journal of experimental & clinical cancer research : CR
影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
Li JJ;Li Q;Du HP;Wang YL;You SJ;Wang F;Xu XS;Cheng J;Cao YJ;Liu CF;Hu LF
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影响因子：
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作者：
Dou C;Wang Y;Li C;Liu Z;Jia Y;Li Q;Yang W;Yao Y;Liu Q;Tu K
通讯作者：
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DOI：
10.1016/j.bbrc.2015.11.132
发表时间：
2016-01-15
影响因子：
3.1
作者：
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通讯作者：
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