A methyltransferase-like 14/miR-99a-5p/tribble 2 positive feedback circuit promotes cancer stem cell persistence and radioresistance via histone deacetylase 2-mediated epigenetic modulation in esophageal squamous cell carcinoma.
A methyltransferase-like 14/miR-99a-5p/tribble 2 positive feedback circuit promotes cancer stem cell persistence and radioresistance via histone deacetylase 2-mediated epigenetic modulation in esophageal squamous cell carcinoma.
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DOI：
10.1002/ctm2.545
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发表时间：
2021-09
影响因子：
10.6
通讯作者：
Zhou Y
中科院分区：
文献类型：
--
作者：
Liu Z;Wu K;Gu S;Wang W;Xie S;Lu T;Li L;Dong C;Wang X;Zhou Y
食管鳞状细胞癌（ESCC）是一种高度侵袭性和耐药的肿瘤。ESCC中的癌症干细胞样细胞（CSC）的生物学意义和分子机制，有助于治疗抗性，如放射抗性，仍然难以捉摸。采用定量真实的实时聚合酶链反应、免疫印迹、免疫组织化学和原位杂交检测食管鳞癌中甲基转移酶样14 miR-99 a-5 p tribble 2（胃癌L14/miR-99 a-5 p/Trib 2）的表达。在体外和体内证实了胃L14/miR-99 a-5 p/TRIB 2的生物学功能。利用质谱分析来鉴定TRIB 2调节的下游蛋白质。采用染色质免疫沉淀（IP）、IP、N6-甲基腺苷（m6 A）-RNA IP、荧光素酶报告基因和泛素化分析来探索这种反馈回路及其下游通路的分子机制。我们发现miR-99 a-5 p在ESCC中显著降低。miR-99 a-5 p抑制CSC的存活和ESCC细胞的放射抗性，miR-99 a-5 p下调预示ESCC患者预后不良。从机制上讲，我们揭示了一个增强CSC特性和ESCC细胞辐射抗性的胃L14-miR-99 a-5 p-TRIB 2正反馈环。胃L14是一种在ESCC中下调的m6 A RNA甲基转移酶，通过靶向其3′非翻译区，通过miR-99 a-5 p介导的TRIB 2 mRNA降解来抑制TRIB 2表达，而TRIB 2则以COP 1依赖性方式诱导泛素介导的胃L14蛋白酶体降解。胃黏膜L14通过调节m6 A介导的DiGeorge critical region 8依赖性pri-mir-99 a加工上调miR-99 a-5 p。TRIB 2的过度激活导致这种积极的电路与辐射抗性和CSC的特点密切相关。此外，TRIB 2激活HDAC 2，随后通过Akt/mTOR/S6 K1信号通路激活诱导p21表观遗传抑制。HDAC 2的药理学抑制在体外和患者来源的异种移植模型中有效减弱TRIB 2介导的作用。我们的数据强调了胃L14/miR-99 a-5 p/TRIB 2轴的存在，并显示其与CSC特征和ESCC的放射抗性正相关，提示了ESCC治疗的潜在治疗靶点。1.胃癌L14通过促进m6 A介导的DGCR 8依赖性pri‐mir‐ 99 a加工上调miR‐ 99 a ‐ 5 p。2. miR-99 a-5 p通过靶向TRIB 2 mRNA的3′ UTR诱导其降解。3. TRIB 2以COP 1依赖性方式诱导泛素介导的胃L14蛋白酶体降解。4.胃癌L14/miR-99 a-5 p/TRIB 2通过诱导依赖于Akt/mTOR/S6 K1通路的HDAC 2介导的p21表观遗传抑制，构成了增强CSC特性和ESCC细胞辐射抗性的正反馈环。
Esophageal squamous cell carcinoma (ESCC) is a highly aggressive and treatment‐resistant tumor. The biological implications and molecular mechanism of cancer stem‐like cells (CSCs) in ESCC, which contribute to therapeutic resistance such as radioresistance, remain elusive. Quantitative real‐time polymerase chain reaction, western blotting, immunohistochemistry, and in situ hybridization assays were used to detect methyltransferase‐like 14 miR‐99a‐5p tribble 2 (METTL14/miR‐99a‐5p/TRIB2) expression in ESCC. The biological functions of METTL14/miR‐99a‐5p/TRIB2 were demonstrated in vitro and in vivo. Mass spectrum analysis was used to identify the downstream proteins regulated by TRIB2. Chromatin immunoprecipitation (IP), IP, N6‐methyladenosine (m6A)‐RNA IP, luciferase reporter, and ubiquitination assays were employed to explore the molecular mechanisms underlying this feedback circuit and its downstream pathways. We found that miR‐99a‐5p was significantly decreased in ESCC. miR‐99a‐5p inhibited CSCs persistence and the radioresistance of ESCC cells, and miR‐99a‐5p downregulation predicted an unfavorable prognosis of ESCC patients. Mechanically, we unveiled a METTL14‐miR‐99a‐5p‐TRIB2 positive feedback loop that enhances CSC properties and radioresistance of ESCC cells. METTL14, an m6A RNA methyltransferase downregulated in ESCC, suppresses TRIB2 expression via miR‐99a‐5p‐mediated degradation of TRIB2 mRNA by targeting its 3′ untranslated region, whereas TRIB2 induces ubiquitin‐mediated proteasomal degradation of METTL14 in a COP1‐dependent manner. METTL14 upregulates miR‐99a‐5p by modulating m6A‐mediated, DiGeorge critical region 8‐dependent pri‐mir‐99a processing. Hyperactivation of TRIB2 resulting from this positive circuit was closely correlated with radioresistance and CSC characteristics. Furthermore, TRIB2 activates HDAC2 and subsequently induces p21 epigenetic repression through Akt/mTOR/S6K1 signaling pathway activation. Pharmacologic inhibition of HDAC2 effectively attenuates the TRIB2‐mediated effect both in vitro and in patient‐derived xenograft models. Our data highlight the presence of the METTL14/miR‐99a‐5p/TRIB2 axis and show that it is positively associated with CSC characteristics and radioresistance of ESCC, suggesting potential therapeutic targets for ESCC treatment. 1. METTL14 upregulates miR‐99a‐5p by promoting m6A‐mediated, DGCR8‐dependent pri‐mir‐99a processing. 2. miR‐99a‐5p induces degradation of TRIB2 mRNA by targeting its 3′ UTR. 3. TRIB2 induces ubiquitin‐mediated proteasomal degradation of METTL14 in a COP1‐dependent manner. 4. METTL14/miR‐99a‐5p/TRIB2 constitutes a positive feedback loop that enhances CSC properties and radioresistance of ESCC cells through inducing HDAC2‐mediated p21 epigenetic repression dependent on Akt/mTOR/S6K1 pathway.
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DOI：
10.1042/bj20141441
发表时间：
2015-04-01
期刊：
The Biochemical journal
影响因子：
--
作者：
Bailey FP;Byrne DP;Oruganty K;Eyers CE;Novotny CJ;Shokat KM;Kannan N;Eyers PA
通讯作者：
Eyers PA
癌症干细胞生态位：在调节肿瘤细胞的干性中，小众的必要性是多么重要？
影响因子：
23.9
作者：
Plaks, Vicki;Kong, Niwen;Werb, Zena
通讯作者：
Werb, Zena
Mettl14通过Notch1的N-6-甲基腺苷抑制膀胱TIC自我更新和膀胱肿瘤发生
影响因子：
37.3
作者：
Gu, Chaohui;Wang, Zhiyu;Tian, Fengyan
通讯作者：
Tian, Fengyan
DOI：
10.1186/s13046-020-01631-w
发表时间：
2020-07-22
影响因子：
11.3
作者：
He, Zhanfeng;Li, Weihao;Zhao, Song
通讯作者：
Zhao, Song
影响因子：
5.1
作者：
Moller, Heidi G.;Rasmussen, Andreas P.;Andersen, Hjalte H.;Johnsen, Kasper B.;Henriksen, Michael;Duroux, Meg
通讯作者：
Duroux, Meg