甲基转移酶样3调节食管鳞癌细胞功能的研究

Study on the function of methyltransferase-like 3 in regulating esophageal squamous cell carcinoma cells

目的:探究甲基转移酶样3(methyltransferase-like 3,METTL3)在食管鳞癌中的表达水平和作用机制。方法:体外构建食管鳞癌细胞Eca-109的METTL3敲低模型,利用实时荧光定量PCR(quantitative real-time PCR,qRTPCR)、Western Blot验证敲低水平,检测肿瘤细胞的增殖、迁移能力。利用差异分析、富集分析以及生存分析等寻找METTL3的靶基因。结果:敲低METTL3可以显著抑制食管癌细胞的增殖和迁移能力;利用生物信息学分析寻找到受METTL3调控的9个可能的靶基因(精脒/精胺N1-乙酰基转移酶1(spermidine/spermine N1-acetyltransferase 1,SAT1)、低密度脂蛋白受体(low density lipoprotein receptor,LDLR)、整合素β1(integrin subunit beta 1,ITGB1)、细胞间黏附分子5(intercellular adhesion molecule 5,ICAM5)、富含谷氨酸的WD重复序列蛋白1(glutamate-rich WD40 repeat containing 1,GRWD1)、含纤维连接蛋白Ⅲ型结构域3A蛋白(fibronectintype-Ⅲdomain-containing protein 3A,FNDC3A)、DnaJ热休克蛋白家族成员C2[DnaJ heat shock protein family(Hsp40)member C2,DNAJC2]、卷曲螺旋域蛋白88C(coiled-coil domain containing 88C,CCDC88C)、Rho GTP酶激活蛋白12(Rho GTPase activating protein 12,ARHGAP12),其中SAT1高表达与食管鳞癌患者预后不良密切相关,差异有统计学意义。结论:METTL3在食管鳞癌中发挥促癌作用,可能通过调控SAT1等靶基因的m6A甲基化影响食管鳞癌患者的预后。

Objective:To explore the expression levels and mechanisms of methyltransferase-like 3(METTL3)in esophageal squamous cell carcinoma.Methods:In vitro construction of METTL3 knockdown model in Eca-109,validated by quantitative real-time PCR(qRT-PCR)and Western Blot to confirm knockdown levels,and assessment of tumor cell proliferation and migration abilities.Exploring METTL3 target genes through differential analysis,enrichment analysis,and survival analysis.Results:Knocking down METTL3 significantly inhibits the proliferation and migration abilities of esophageal cancer cells.Through bioinformatics analysis,we identified 9 potential target genes regulated by METTL3 spermidine/spermine N1-acetyltransferase 1(SAT1),low density lipoprotein receptor(LDLR),integrin subunit beta 1(ITGB1),intercellular adhesion molecule 5(ICAM5),glutamate-rich WD40 repeat containing 1(GRWD1),fibronectintype-Ⅲdomain-containing protein 3A(FNDC3A),DnaJ heat shock protein family(Hsp40)member C2(DNAJC2),coiled-coil domain containing 88C(CCDC88C),Rho GTPase activating protein 12(ARHGAP12).High expression of SAT1 is closely associated with poor prognosis in esophageal squamous cell carcinoma patients.Conclusions:METTL3 exerts a pro-cancer role in esophageal squamous cell carcinoma,potentially impacting the prognosis of esophageal squamous cell carcinoma patients by regulating m6A methylation of target genes such as SAT1.