基于METTL14/ERα信号探讨m6A调控子宫内膜癌转移的机制研究  

Exploration on the mechanism of m6A on regulating metastasis of endometrial cancer based on METTL14/ERαsignaling

作　　者：欧阳诗洁 熊琼英 李仙丽[1] OUYANG Shijie;XIONG Qiongying;LI Xianli(Xiaogan Hospital Affiliated to Wuhan University of Science and Technology,Xiaogan,Hubei 432000,China)

机构地区：[1]武汉科技大学附属孝感医院,湖北孝感432000

出　　处：《中国优生与遗传杂志》2023年第3期439-446,共8页Chinese Journal of Birth Health & Heredity

摘　　要：目的探讨甲基转移酶样因子14(METTL14)失调引起的异常N6-甲基腺苷(m6A)修饰在子宫内膜癌(EC)侵袭和转移中的作用。方法收集96名2017—2021年在武汉科技大学附属孝感医院接受治疗性手术的EC患者。从冷冻组织中分离RNA(70对)或蛋白质(10对),用于定量实时PCR(qPCR)或免疫印迹分析,以评估METTL14在EC中的表达。评估METTL14的表达及其与EC临床病理学特征的相关性。在体外和体内测定了METTL14在EC中的生物学效应。将甲基化RNA免疫沉淀测序(MeRIP-seq)与RNA测序(RNA-seq)相结合,并在m6A斑点印迹之后,采用MeRIP-qPCR、RIP-qPCR或双荧光素酶报告基因分析来筛选和验证METTL14的候选靶标。结果与匹配的相邻组织相比,EC中METTL14的mRNA和蛋白质水平显著下调。与METTL14高表达组相比,METTL14低表达组FIGO分期、浸入深度、淋巴血管侵犯、淋巴结转移、肿瘤转移例数显著增加(P<0.05)。在功能上,METTL14在体外和体内抑制EC细胞的增殖和侵袭能力。从机制上讲,METTL14介导的m6A去甲基化导致含有雌激素受体α(ERα)的转录后抑制。此外,与METTL14相比,ERα诱导肿瘤的致癌行为。结论METTL14通过m6A依赖性方式在EC细胞中减弱ERα的表达,进而抑制肿瘤转移和侵袭。Objective To explore the effects of abnormal N6-methyladenosine(m6A)modification induced by imbalance of methyltransferase-like factor 14(METTL14)on invasion and metastasis of endometrial cancer(EC).Methods A total of 96 patients with EC undergoing surgical treatment Xiaogan Hospital Affiliated to Wuhan University of Science and Technology were enrolled between 2017 and 2021.RNA(70 pairs)or proteins(10 pairs)was isolated from frozen tissues for real-time quantitative PCR(qPCR)or Western blot analysis so as to assess the expression of METTL14 in EC.The expression of METTL14 and its correlation with clinicopathological characteristics of EC were evaluated.The biological effects of METTL14 in EC were detected in vitro and in vivo.The methylated RNA immunoprecipitation-sequencing(MeRIP-seq)was combined with RNA-sequencing(RNA-seq).After m6A dot blot,candidate targets of METTL14 were screened out and validated by MeRIP-qPCR,RIP-qPCR,or dual-luciferase reporter assay.Results Compared with matched adjacent tissues,levels of METTL14 mRNA and protein were significantly down-regulated in EC.Compared with high-expression METTL14 group,FIGO staging,infiltration depth,number of cases with lymphovascular invasion,lymph node metastasis and tumor metastasis were significantly increased in low-expression group(P<0.05).In terms of function,METTL14 could inhibit the proliferation and invasion of EC cells in vitro and in vivo.In terms of mechanism,METTL14-mediated demethylation of m6A could cause post-transcription inhibition of estrogen receptorα(ERα).Furthermore,ERαcould induce oncogenic behaviors of tumors compared with METTL14.Conclusion METTL14 could reduce the expression of ERαin EC cells,showing m6A dependence,and thereby inhibiting metastasis and invasion of tumors.

关 键 词：甲基转移酶样因子14 N6-甲基腺苷 子宫内膜癌 雌激素受体Α 

分 类 号：R737.33[医药卫生—肿瘤]