赖氨酰氧化酶LOXL4去组蛋白H4K8乙酰化促进结直肠癌进展的分子机制研究
项目来源
国（略）科（略）(（略）C（略）
项目主持人
吴（略）
项目受资助机构
苏（略）
项目编号
3（略）1（略）
立项年度
2（略）
立项时间
未（略）
研究期限
未（略） （略）
项目级别
国（略）
受资助金额
2（略）0（略）
学科
生（略）-（略）与（略）息（略）传（略）构（略）
学科代码
C（略）6（略）6（略）
基金类别
青（略）基（略）
关键词
L（略）4（略）人（略） （略）去（略） （略）直（略）;（略）K（略）化（略）L（略）4（略）人（略） （略）去（略） （略）直（略）;（略）K（略）化
参与者
张（略）黄（略）徐（略）李（略）郑（略）胡（略）
参与机构
苏（略） ；同济大学
项目标书摘要：表观（略）发展中起重要作用。（略）在结直肠癌中高表达（略）能。我们预实验发现（略）异降低组蛋白H4K（略）细胞增殖、侵袭和迁（略）在体外去除H4K8（略）说:LOXL4是一（略），通过去除H4K8（略）进结肠癌进展。围绕（略）质谱技术、基因组学（略）模型等手段1研究L（略）乙酰化的作用机理;（略）因;3阐明LOXL（略）调控靶基因表达促进（略）。本项目的顺利实施（略）酰化酶活性，发现结（略）为针对结直肠癌靶向（略）依据。
Applicati（略）: Epigene（略）ions are （略）ents in c（略）ancer(CRC（略）dase like（略）s been re（略）e overexp（略）RC,which （略）lies its （略）otential.（略）nary data（略）t,histone（略）lation wa（略）reduced w（略）as ectopi（略）ssed in H（略）s.In vitr（略）4K8 deace（略）s also ob（略）ncubating（略）urified L（略）re histon（略）ore,in HC（略）ctal canc（略）e,overexp（略）LOXL4 sig（略）increased（略）feration,（略）nd invasi（略）e,we prop（略）XL4 is a （略）ne deacet（略）accelerat（略）al cancer（略）n by deac（略）istone H4（略）g mass sp（略）technique（略）e genomic（略）nd mouse （略）odel,this（略）ll furthe（略）r the mec（略）OXL4 cata（略）tylation （略）dentify L（略） genes;3)（略）he mechan（略）4 in prom（略）ectal can（略）sion by r（略）tone H4K8（略）n.The pro（略） would po（略）ead to id（略）vel histo（略）ase and r（略） drug tar（略）rectal ca（略）
项目受资助省
江（略）
项目结题报告(全文)
LOXL4(Lys（略） like 4)是（略）l oxidase（略）研究发现该家族蛋白（略）型赖氨酰氧化酶，通（略）原蛋白赖氨酰氧化反（略）节细胞外基质的稳态（略）家族在肿瘤的增殖、（略）要作用,LOXL4（略）尚不明确。组蛋白修（略）方面，并且与结直肠（略）直肠癌细胞系和肿瘤（略）H4乙酰化水平明显（略）化酶抑制剂作为结直（略）临床前或临床研究阶（略）被认为是有希望的结（略）利用TCGA(Th（略）nom Eatla（略） Expressi（略）数据库分析，发现相（略）肿瘤组织中的LOX（略）LOXL4表达更高（略）差。利用构建过表达（略）直肠癌细胞系进行的（略）Trans-Wel（略）实验分别在细胞水平（略）L4会促进结直肠癌（略）免疫印迹和Orbi（略）技术发现LOXL4（略）4八号赖氨酸(H4（略）NA-Seq和Ch（略）)-Seq发现LO（略）4K8的乙酰化水平（略）atenin通路相（略）次鉴定出LOXL4（略）性去乙酰化酶，通过（略）调控WNT/β-c（略）因的转录促进结直肠（略）子机制，有望为找寻（略）生物标志物提供新思（略）下，我们还发现赖氨（略）够发生N-豆蔻酰化（略）过介导疏水相互作用（略）相分离，招募并活化（略）性进展的分子机制。（略）靶点，为靶向EZH（略）。
1. Nominal unification with atom-variables.J.Symb.Comput.,90:42-64
2.Emerging role of SIRT2 in non-small cell lung cancer
- 关键词：
- sirtuin 2; NSCLC; function; pathogenic mechanism; therapeutic target;TUMOR-SUPPRESSOR; SIRT2-MEDIATED DEACETYLATION; HISTONE DEACETYLASE;ACETYLATION; SIRTUINS; PROTEIN; DEHYDROGENASE; GENE; PROMOTES;TUMORIGENESIS
Non-small cell lung cancer (NSCLC) is one of the most devastating cancer types, accounting for >80% of lung cancer cases. The median relative survival time of patients with NSCLC is <1 year. Lysine acetylation is a major post-translational modification that is required for various biological processes, and abnormal protein acetylation is associated with various diseases, including NSCLC. Protein deacetylases are currently considered cancer permissive partly due to malignant cells being sensitive to deacetylase inhibition. Sirtuin 2 (SIRT2), a primarily cytosolic nicotinamide adenine dinucleotide-dependent class III protein deacetylase, has been shown to catalyze the removal of acetyl groups from a wide range of proteins, including tubulin, ribonucleotide reductase regulatory subunit M2 and glucose-6-phosphate dehydrogenase. In addition, SIRT2 is also known to possess lysine fatty deacylation activity. Physiologically, SIRT2 serves as a regulator of the cell cycle and of cellular metabolism. It has been shown to play important roles in proliferation, migration and invasion during carcinogenesis. It is notable that both oncogenic and tumor suppressive functions of SIRT2 have been described in NSCLC and other cancer types, suggesting a context-specific role of SIRT2 in cancer progression. In addition, inhibition of SIRT2 exhibits a broad anticancer effect, indicating its potential as a therapeutic for NSCLC tumors with high expression of SIRT2. However, due to the diverse molecular and genetic characteristics of NSCLC, the context-specific function of SIRT2 remains to be determined. The current review investigated the functions of SIRT2 during NSCLC progression with regard to its regulation of metabolism, stem cell-like features and autophagy.
...3.赖氨酰氧化酶LOXL4去组蛋白H4K8乙酰化促进结直肠癌进展的分子机制研究结题报告
- 吴孟；
- 《苏州大学；》
- 2021年
- 报告
LOXL4(Lysyl oxidases like 4)是赖氨酰氧化酶(Lysyl oxidases,LOX)家族成员，研究发现该家族蛋白是一类铜离子依赖的分泌型赖氨酰氧化酶，通过催化弹性纤维蛋白和胶原蛋白赖氨酰氧化反应使之相互交联，参与调节细胞外基质的稳态。近年有研究揭示LOX家族在肿瘤的增殖、侵袭和迁移过程中发挥重要作用,LOXL4在结直肠癌中发挥的作用尚不明确。组蛋白修饰是表观遗传调控的重要方面，并且与结直肠癌发生发展密切相关，结直肠癌细胞系和肿瘤组织中总体组蛋白H3和H4乙酰化水平明显下降，多个组蛋白去乙酰化酶抑制剂作为结直肠癌靶向治疗药物正处于临床前或临床研究阶段，组蛋白去乙酰化酶也被认为是有希望的结直肠癌治疗靶点。本项目利用TCGA(The Cancer Genom Eatlas)和GEO(Gene Expression Omnibus)数据库分析，发现相比癌旁组织结直肠癌病人肿瘤组织中的LOXL4的表达水平更高，且LOXL4表达更高的结直肠癌患者的预后更差。利用构建过表达或者敲除LOXL4的结直肠癌细胞系进行的CCK8细胞增殖实验和Trans-Well以及裸鼠移植瘤模型等实验分别在细胞水平和动物水平上发现LOXL4会促进结直肠癌的发生发展。在我们通过免疫印迹和Orbitrap LC-MS等技术发现LOXL4可以特异性去除组蛋白H4八号赖氨酸(H4K8)上的乙酰基团。RNA-Seq和ChIP(染色质免疫共沉淀)-Seq发现LOXL4通过影响组蛋白H4K8的乙酰化水平从而调控WNT/β-catenin通路相关基因的转录。本研究首次鉴定出LOXL4是组蛋白H4K8的特异性去乙酰化酶，通过去乙酰化组蛋白H4K8调控WNT/β-catenin通路相关基因的转录促进结直肠癌增殖、迁移和侵袭的分子机制，有望为找寻结直肠癌预防和治疗新的生物标志物提供新思路。此外，在本项目资助下，我们还发现赖氨酸甲基转移酶EZH2能够发生N-豆蔻酰化修饰，并阐明豆蔻酰化通过介导疏水相互作用驱动EZH2发生液—液相分离，招募并活化STAT3,促进肺癌恶性进展的分子机制。为肺癌的精准治疗提供新靶点，为靶向EZH2的药物开发提供新视角。
...4.Myristoylation-mediated phase separation of EZH2 compartmentalizes STAT3 to promote lung cancer growth
- 《CANCER LETTERS》
- 2021年
- 516卷
- 期
- 期刊
N-myristoylation is a crucial signaling and pathogenic modification process that confers hydrophobicity to cytosolic proteins. Although different large-scale approaches have been applied, a large proportion of myristoylated proteins remain to be identified. EZH2 is overexpressed in lung cancer cells and exerts oncogenic effects via its intrinsic methyltransferase activity. Using a well-established click chemistry approach, we found that EZH2 can be modified by myristoylation at its N-terminal glycine in lung cancer cells. Hydrophobic interaction is one of the main forces driving or stabilizing liquid-liquid phase separation (LLPS), raising the possibility that myristoylation can modulate LLPS by mediating hydrophobic interactions. Indeed, myristoylation facilitates EZH2 to form phase-separated liquid droplets in lung cancer cells and in vitro. Furthermore, we provide evidence that myristoylation-mediated LLPS of EZH2 compartmentalizes its non-canonical substrate, STAT3, and activates STAT3 signaling, ultimately resulting in accelerated lung cancer cell growth. Thus, targeting EZH2 myristoylation may have significant therapeutic efficacy in the treatment of lung cancer. Altogether, these observations not only extend the list of myristoylated proteins, but also indicate that hydrophobic lipidation may serve as a novel incentive to induce or maintain LLPS.
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