摘要
目的探究甲基转移酶样蛋白3(methyltransferase-like 3,METTL3)对子宫内膜蜕膜化的作用。方法采用生物信息学分析26个N6-甲基腺嘌呤(N6-methyladenosine,m6A)调节因子在反复种植失败(recurrent implantation failure,RIF)、复发性流产(recurrent spontaneous abortion,RSA)及子宫内膜基质细胞蜕膜化中的表达情况;使用醋酸甲羟孕酮和二丁酰环磷酸腺苷诱导人子宫内膜基质细胞(human endometrial stromal cells,HESCs)发生蜕膜化,检测对照组和蜕膜化组中的总m6A水平、METTL3、叉头框蛋白O1(Forkhead box protein O1,FOXO1)及蜕膜化标志物胰岛素样生长因子结合蛋白1(insulin-like growth factor binding protein 1,IGFBP1)和催乳素的表达水平;通过慢病毒构建METTL3干扰的HESCs模型,CCK-8及Edu检测阴性对照组和敲低METTL3组的增殖情况。慢病毒转染成功后再诱导蜕膜化,检测FOXO1及蜕膜化标志物的表达变化。结果生物信息学分析结果显示,METTL3在RIF患者和RSA患者子宫内膜中的表达显著下调(P=0.014,P=0.016),而在蜕膜化的子宫内膜中显著上调(P=0.029)。体外诱导HESCs蜕膜化,蜕膜化组中总m6A、METTL3、FOXO1蛋白表达量升高(P=0.015,P=0.016,P=0.004)。敲低METTL3后HESCs中FOXO1蛋白表达量降低(P=0.009);CCK-8及Edu结果显示,敲低METTL3后HESCs的增殖被抑制。体外诱导敲低METTL3的HESCs蜕膜化后,IGFBP1 mRNA、PRL mRNA、FOXO1蛋白表达量较阴性对照组差异均无统计学意义(均P>0.05)。结论在子宫内膜基质细胞中,METTL3能够调控FOXO1的表达,促进IGFBP1、催乳素的表达,影响子宫内膜细胞蜕膜化。
Objective:To explore the role of methyltransferase-like 3(METTL3)on endometrial decidualization.Methods:Bioinformatics was used to analyze the expression of 26 N6-methyladenosine(m6A)regulators in recurrent implantation failure(RIF),recurrent spontaneous abortion(RSA)and the decidualization of endometrial stromal cells.Using Medroxyprogesterone and Dibutyryl-cAMP to induce human endometrial stromal cell s(HESCs)decidualization,we examined changes in the expression of total m6A,METTL3,Forkhead framing protein O1(FOXO1),and decidualization markers insulin-like growth factor binding protein 1(IGFBP1)and prolactin in the control and decidualization group.Constructing HESC models with METTL3 interference using lentivirus,we used CCK-8 and Edu to detect proliferation in the negative control and knockdown METTL3 group.Lentiviral transfection was successfully followed by induction of decidualization,and we examined changes in the expression of FOXO1 and decidualization markers.Results:The results of bioinformatics analysis showed that the expression of METTL3 was significantly downregulated in the endometrium of patients with RIF and RSA(P=0.014,P=0.016),while it was significantly upregulated in the endometrium with decidualization(P=0.029).In vitro,induction of HESCs decidualization increased in the expression levels of total m6A,METTL3,and FOXO1 proteins in the decidualization group(P=0.015,P=0.016,P=0.004).Knocking down METTL3 resulted in a decrease in FOXO1 protein expression in HESCs(P=0.009).The CCK-8 and Edu results showed that the proliferation of HESCs was inhibited after knocking down METTL3.After inducing the knockdown of METTL3 in vitro,there was no statistically significant difference in the expression levels of IGFBP1 mRNA,PRL mRNA,and FOXO1 protein in HESCs compared with the negative control group(all P>0.05).Conclusion:In endometrial stromal cells,METTL3 can regulate the expression of FOXO1,promote the expression of IGFBP1 and PRL,and affect the decidualization of endometrial cells.
作者
葛挺
张曼丽
张于念
徐强
腊晓琳
Ge Ting;Zhang Manli;Zhang Yunian;Xu Qiang;La Xiaolin(Center for Reproductive Medicine,First Affiliated Hospital of Xinjiang Medical University,Urumqi 830000,China;Research Center for Comprehensive Prevention and Treatment of Reproductive Diseases and Birth Defects,Urumgi 830000,China)
出处
《中华生殖与避孕杂志》
CAS
CSCD
北大核心
2024年第11期1146-1155,共10页
Chinese Journal of Reproduction and Contraception
基金
国家自然科学基金地区项目(81960289)
新疆维吾尔自治区自然科学基金青年基金(2021D01C342)。
关键词
N6-甲基腺嘌呤
甲基转移酶样蛋白3
蜕膜化
反复种植失败
复发性流产
N6-methyladenosine
Methyltransferase-like 3
Decidualization
Recurrent implantation failure
Recurrent spontaneous abortion
