METTL14-mediated epitranscriptome modification of MN1 mRNA promote tumorigenicity and all-trans-retinoic acid resistance in osteosarcoma.
METTL14-mediated epitranscriptome modification of MN1 mRNA promote tumorigenicity and all-trans-retinoic acid resistance in osteosarcoma.
复制标题
DOI：
10.1016/j.ebiom.2022.104142
复制
发表时间：
2022-08
期刊：
影响因子：
11.1
通讯作者：
Xie, Xian-Biao
中科院分区：
文献类型：
--
作者：
Li, Hong -Bo;Huang, Gang;Tu, Jian;Lv, Dong-Ming;Jin, Qing-Lin;Chen, Jun -Kai;Zou, Yu -Tong;Lee, Dung -Fang;Shen, Jing-Nan;Xie, Xian-Biao
骨肉瘤是青少年最常见的原发性恶性骨肿瘤。OS进展和转移背后的分子机制仍然知之甚少，这限制了当前治疗的有效性。RNA N6-甲基腺苷（m6 A）修饰在影响RNA命运中起关键作用。然而，m6 A修饰的生物学意义及其在OS发生中的潜在调控机制仍不清楚。采用液相色谱-串联质谱法（LC-MS/MS）、斑点印迹法和比色ELISA法检测m6 A水平。采用Western blotting、实时荧光定量PCR（RT-qPCR）和免疫组织化学（IHC）检测胃L14的表达水平。采用甲基化RNA免疫沉淀测序（MeRIP-seq）和转录组RNA测序（RNA-seq）方法筛选目的基因。进行RNA下拉和RNA免疫沉淀（RIP）测定以探索靶基因和相关m6 A“阅读器”的特异性结合。使用RNA稳定性和多聚核糖体分析测定来检测CITL 14下游基因的半衰期和翻译效率。通过免疫组化和临床资料分析，探讨了胃L14及其下游靶基因与OS预后的临床相关性。我们观察了OS中丰富的m6 A修饰，揭示了胃L14在促进OS进展中起致癌作用。MeRIP-seq和RNA-seq分析表明MN 1是胃癌L14的下游基因。MN 1与肿瘤进展和OS中全反式维甲酸（ATRA）化疗耐药性有关。从机制上讲，MN 1被胃L14甲基化，特别是在编码序列（CDS）区域，并且这种修饰被特异性m6 A阅读器胰岛素样生长因子2 mRNA结合蛋白2（IGF 2BP 2）识别，以防止MN 1 mRNA降解并促进其翻译效率。免疫组化显示MN 1与胃L14、IGF 2BP 2在OS组织中的表达呈正相关。与这些分子中的任何一种相比，胃L14-IGF 2BP 2-MN 1组显示出对OS患者更有希望的预后价值。我们的研究表明，胃L14作为m6 A RNA甲基化酶，通过调节MN 1的稳定性和翻译效率，促进OS进展和ATRA耐药，从而为OS患者的预后预测提供了基础生物标志物。本工作得到了国家自然科学基金项目（赠款81972510和81772864）的资助。
Osteosarcoma (OS) is the most common primary malignant bone tumor in adolescents. The molecular mechanism behind OS progression and metastasis remains poorly understood, which limits the effectiveness of current therapies. RNA N6-methyladenosine (m6A) modification plays a critical role in influencing RNA fate. However, the biological significance of m6A modification and its potential regulatory mechanisms in the development of OS remain unclear. Liquid chromatography-tandem mass spectrometry (LC-MS/MS), dot blotting, and colorimetric ELISA were used to detect m6A levels. Western blotting, quantitative real-time PCR (RT-qPCR) and immunohistochemistry (IHC) were used to investigate METTL14 expression levels. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) and transcriptomic RNA sequencing (RNA-seq) were used to screen the target genes of METTL14. RNA pull-down and RNA immunoprecipitation (RIP) assays were conducted to explore the specific binding of target genes and relevant m6A “readers”. RNA stability and polysome analysis assays were used to detect the half-lives and translation efficiencies of the downstream genes of METTL14. IHC and clinical data were applied to explore the clinical correlations of METTL14 and its downstream target genes with the prognosis of OS. We observed the abundance of m6A modifications in OS and revealed that METTL14 plays an oncogenic role in facilitating OS progression. MeRIP-seq and RNA-seq revealed that MN1 is a downstream gene of METTL14. MN1 contributes to tumor progression and all-trans-retinoic acid (ATRA) chemotherapy resistance in OS. Mechanistically, MN1 is methylated by METTL14, specifically in the coding sequence (CDS) regions, and this modification is recognized by the specific m6A reader insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) to prevent MN1 mRNA degradation and promote it translation efficiency. IHC showed that MN1 expression was positively correlated with METTL14 and IGF2BP2 expression in OS tissues. The METTL14-IGF2BP2-MN1 panel demonstrated more promising prognostic value for OS patients than any of these molecules individually. Our study revealed that METTL14 contributes to OS progression and ATRA resistance as an m6A RNA methylase by regulating the stability and translation efficiency of MN1 and thus provides both an underlying biomarker panel for prognosis prediction in OS patients. This work was supported by the National Natural Science Foundation of China (Grants 81972510 and 81772864).
登录
查看更多内容
影响因子：
2.4
作者：
Barroga, EF;Kadosawa, T;Fujinaga, T
通讯作者：
Fujinaga, T
影响因子：
11.4
作者：
Carella, C.;Bonten, J.;Grosveld, G. C.
通讯作者：
Grosveld, G. C.
影响因子：
50.3
作者：
Li Z;Weng H;Su R;Weng X;Zuo Z;Li C;Huang H;Nachtergaele S;Dong L;Hu C;Qin X;Tang L;Wang Y;Hong GM;Huang H;Wang X;Chen P;Gurbuxani S;Arnovitz S;Li Y;Li S;Strong J;Neilly MB;Larson RA;Jiang X;Zhang P;Jin J;He C;Chen J
通讯作者：
Chen J
影响因子：
20.3
作者：
Heuser, Michael;Argiropoulos, Bob;Humphries, R. Keith
通讯作者：
Humphries, R. Keith
DOI：
10.1038/nrc.2017.23
发表时间：
2017-06
期刊：
Nature reviews. Cancer
影响因子：
--
作者：
Jiang Q;Crews LA;Holm F;Jamieson CHM
通讯作者：
Jamieson CHM