METTL3 regulates m6A in endometrioid epithelial ovarian cancer independently of METTl14 and WTAP
METTL3 regulates m6A in endometrioid epithelial ovarian cancer independently of METTl14 and WTAP
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DOI：
10.1002/cbin.11459
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发表时间：
2020-09-11
影响因子：
3.9
通讯作者：
Zuo, Ying
中科院分区：
文献类型：
--
作者：
Ma, Zhao;Li, Qin;Zuo, Ying
N6-甲基腺苷（m6 A）RNA甲基化是一种常见的RNA修饰，已被确定在肿瘤发生和癌症发展中起关键作用。包括甲基转移酶样3（甲基转移酶L3）、甲基转移酶L14和肾母细胞瘤1相关蛋白（Wilms tumor 1-associated protein，WTAP）的m6 A“写入者”有助于m6 A修饰过程的启动。然而，m6 A甲基转移酶复合物的协调在类上皮性卵巢癌（EEOC）中尚未完全了解。本研究采用定量聚合酶链反应（qPCR）、免疫组化和western blot分析方法检测了33例EEOC中胃L3、胃L14和WTAP的mRNA和蛋白水平。通过点图检测总体m6 A甲基化。EEOC组织中的胃L3表达和总m6 A水平升高，而胃L14和WTAP的表达与邻近组织相比无显著差异。胃L3表达是EEOC恶性程度低、生存期长的独立相关因素。此外，使用CCK-8测定、transwell测定、伤口愈合测定和TUNEL测定，与阴性对照和WTAP或WTL 14敲除的细胞相比，TOV-112 D和CRL-11731 D细胞中WTL 3敲除削弱了细胞增殖和迁移的能力，并促进了细胞凋亡。此外，通过RIP-qPCR测定，胃L3敲低还减少了TOV-112 D和CRL-11731 D细胞中与卵巢癌相关的基因（包括EIF 3C、AXL、CSF-1、FZD 10）的m6 A富集。总之，高表达的胃L3通过调节异常m6 A RNA甲基化指示EEOC的低恶性度和存活。不依赖于WTAP和胃L14，胃L3介导的m6 A修饰被认为是m6 A调节的新机制和EEOC的潜在治疗靶点。
N6-methyladenosine (m6A) RNA methylation, one of the common RNA modifications, has been determined to execute crucial functions in tumorigenesis and cancer development. The m6A "writers" including methyltransferase like 3 (METTL3), METTL14, and Wilms tumor 1-associated protein (WTAP) contribute to the m6A modification process initiation. However, the coordination of m6A methyltransferase complex is not fully understood in endometrioid epithelial ovarian cancer (EEOC). In this study, mRNA and protein levels of METTL3, METTL14, and WTAP were detected in 33 EEOC cases using quantitative polymerase chain reaction (qPCR), immunohistochemistry, and western blot analysis. The overall m6A methylation was detected by dot plot. The METTL3 expression and overall m6A level were elevated in EEOC tissues, while the expressions of METTL14 and WTAP have no significant difference in EEOC compared to the adjacent tissues. The expression of METTL3 was an independent factor that correlated with poor malignancy and survival of EEOC patients. Moreover, METTL3 knockdown in TOV-112D and CRL-11731D cells weakened the capability of cell proliferation and migration, and promoted cell apoptosis compared to negative control and cells with WTAP or METTL14 knockdown using CCK-8 assay, transwell assay, wound healing assay, and TUNEL assay. Furthermore, METTL3 knockdown also reduced m6A enrichment of the genes associated with ovarian cancer including EIF3C, AXL, CSF-1, FZD10 in TOV-112D, and CRL-11731D cells by RIP-qPCR assay. Taken together, the high expressed METTL3 indicated poor malignancy and survival of EEOC via modulating the aberrant m6A RNA methylation. METTL3-mediated m6A modification, independent of WTAP and METTL14, was considered as a novel mechanism underlying m6A modulation and a potential therapeutic target of EEOC.