METTL3/DUXAP8轴促进涎腺腺样囊性癌细胞的增殖、迁移和侵袭  

METTL3/DUXAP8 axis promotes proliferation,migration and invasion of salivary adenoid cystic carcinoma cells

作　　者：赵琦 高万鹏 王嘉乐 刘荣 史明锐 任骋昊 杨子桧 白朕卿 杨新杰 ZHAO Qi;GAO Wanpeng;WANG Jiale;LIU Rong;SHI Mingrui;REN Chenghao;YANG Zihui;BAI Zhenqing;YANG Xinjie(College of Life Sciences,Yan'an University,Shaanxi Province,China,716000;State Key Laboratory of Oral&Maxillofacial Reconstruction and Regeneration,National Clinical Research Center for Oral Diseases,Shaanxi Clinical Research Center for Oral Diseases,Department of Oral and Maxillofacial Surgery,The Third Hospital Affiliated of Air Force Military Medical University,Xi'an)

机构地区：[1]延安大学生命科学学院,716000 [2]口颌系统重建与再生全国重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔疾病临床医学研究中心,空军军医大学第三附属医院口腔颌面外科

出　　处：《实用口腔医学杂志》2024年第3期337-343,共7页Journal of Practical Stomatology

基　　金：国家自然科学基金(编号:82173165);国家自然科学基金青年项目(编号:82002867);国家口腔疾病临床医学研究中心专项课题(编号:LCB202008)。

摘　　要：目的:研究甲基转移酶样3(METTL3)介导的m6A修饰调控双同源盒A假基因8(DUXAP 8)对涎腺腺样囊性癌细胞SACC-LM的增殖、迁移和侵袭能力的影响及其潜在的分子机制。方法:通过肿瘤组织和癌旁组织全转录组测序筛选出差异基因DUXAP 8并进行qRT-PCR验证;采用m6A修饰位点预测网站SRAMP预测DUXAP 8上的m6A修饰位点;qRT-PCR、Western blot检测m6A修饰和上皮-间质转化(EMT)相关基因mRNA及蛋白水平。干扰或过表达METTL3和DUXAP 8,通过CCK-8、划痕、侵袭实验检测各组细胞的增殖、迁移和侵袭能力;结合MeRIP-qPCR检测METTL3和DUXAP 8的相关性。结果:DUXAP 8在SACC肿瘤组织的表达显著高于癌旁组织(P<0.05);干扰DUXAP 8可以显著抑制SACC-LM细胞的增殖、迁移和侵袭能力以及EMT相关基因表达水平(P<0.05);DUXAP 8上存在多个可信度较高的m6A修饰位点;METTL3在肿瘤组织高表达且较其他相关基因差异最为显著(P<0.05);METTL3作为甲基转移酶调控DUXAP 8的表达;下调METTL3可以显著抑制SACC-LM细胞的增殖、迁移和侵袭能力,并可部分逆转DUXAP 8过表达对这些能力的促进作用(P<0.05)。结论:METTL3介导的m6A修饰上调DUXAP 8的表达,从而促进了SACC细胞的增殖、迁移和侵袭能力。Objective:To investigate the effects of methyltransferases like 3(METTL3)mediated m6A modification of double homology cassette A pseudogene 8(DUXAP 8)on the proliferation,migration and invasion of salivary adenoid cystic carcinoma SACC-LM cells and its potential molecular mechanisms.Methods:Whole-transcriptome sequencing showed that DUXAP 8 was highly expressed in SACC than in para-cancerous tissues(P<0.05).The m6A modification sites on DUXAP 8 were predicted using the SRAMP website,and the mRNA and protein expression of m6A-modified genes and the genes associated with the epithelial-mesenchymal transition(EMT)was measured by qRT-PCR and Western blot,respectively.METTL3 and DUXAP 8 was knocked down or overexpressed in SACC-LM cells,and the proliferation,migration,and invasion of the cells were assessed by CCK-8,scratch and Transwell assays.The correlation between METTL3 and DUXAP 8 was evaluated using MeRIP-qPCR.Results:The expression of DUXAP 8 in SACC tumor was higher than that in para-cancerous tissues(P<0.05).Knockdown of DUX AP8 reduced proliferation,migration and invasion of SACC-LM cells,as well as the expression of EMT-related genes(P<0.05).Multiple m6A modification sites of high confidenc e were found on DUXAP 8.METTL3 was highly expressed in tumor tissues,more than other related genes(P<0.05)and enzyme-encoding genes in SACC-LM cells(P<0.05).METTL3 was found to function as a methyltransferase to regulate the expression of DUXAP 8,and downregulation of METTL3 inhibited proliferation,migration and invasion of SACC-LM cells and partially reversed the promotion of these activities induced by DUXAP 8 overexpression(P<0.05).Conclusion:METTL3-medi ated m6A modification upregulated DUXAP 8 expression,which promotes the proliferation,migration and invasion of SACC cells.

关 键 词：涎腺腺样囊性癌 METTL3 DUXAP 8 增殖 侵袭 转移 

分 类 号：R739.8[医药卫生—肿瘤]