Zhou Hongyu, Zhang Nana
Department of Otorhinolaryngology Head and Neck Surgery, Wuhan Fourth Hospital, Wuhan 430033, Hubei, China.
Open Med (Wars). 2022 Jul 12;17(1):1241-1251. doi: 10.1515/med-2022-0515. eCollection 2022.
This study was conducted to investigate the effect of microRNA-212-5p (miR-212-5p) on the proliferation and apoptosis of nasopharyngeal carcinoma (NPC) cells. Microarray datasets (EXP00394 and EXP00660) were downloaded from the dbDEMC database, and the differentially expressed microRNAs between high-grade and low-grade NPC were analyzed. miR-212-5p and methyltransferase like 3 (METTL3) expression levels in NPC tissues and cells were determined by the quantitative real-time polymerase chain reaction and Western blot. Besides, the relationship between miR-212-5p expression and clinicopathological characteristics of patients was analyzed by the Chi-square test. Cell counting kit-8 assay, 5-ethynyl-2-deoxyuridine (EdU) assay, and flow cytometry were adopted to detect the effect of miR-212-5p on the cell proliferation and apoptosis. Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analysis were performed to explore the potential biological functions and the signal pathways related to the target genes of miR-212-5p. Bioinformatics prediction and dual luciferase reporter gene assay were used to verify the relationship between miR-212-5p and METTL3 3' untranslated region. Besides, western blot was adopted to detect the expression of METTL3. Gene set enrichment analysis was performed to analyze the downstream pathways in which METTL3 was enriched. It was found that miR-212-5p was downregulated in NPC tissues, and the low miR-212-5p expression was associated with lymph node metastasis and poor differentiation. miR-212-5p overexpression inhibited the growth and promoted apoptosis of NPC cells; miR-212-5p inhibition functioned oppositely. Mechanistically, miR-212-5p inhibited the proliferation and promoted apoptosis of NPC cells via suppressing METTL3 expression. miR-212-5p/METTL3 was associated with processes of RNA transport and cell cycle. In conclusion, miR-212-5p inhibits the progression of NPC by targeting METTL3.
本研究旨在探讨微小RNA-212-5p（miR-212-5p）对鼻咽癌（NPC）细胞增殖和凋亡的影响。从dbDEMC数据库下载微阵列数据集（EXP00394和EXP00660），并分析高级别和低级别NPC之间差异表达的微小RNA。通过定量实时聚合酶链反应和蛋白质免疫印迹法测定NPC组织和细胞中miR-212-5p和甲基转移酶样3（METTL3）的表达水平。此外，采用卡方检验分析miR-212-5p表达与患者临床病理特征之间的关系。采用细胞计数试剂盒-8法、5-乙炔基-2'-脱氧尿苷（EdU）法和流式细胞术检测miR-212-5p对细胞增殖和凋亡的影响。进行京都基因与基因组百科全书和基因本体分析，以探索与miR-212-5p靶基因相关的潜在生物学功能和信号通路。利用生物信息学预测和双荧光素酶报告基因测定法验证miR-212-5p与METTL3 3'非翻译区之间的关系。此外，采用蛋白质免疫印迹法检测METTL3的表达。进行基因集富集分析，以分析METTL3富集的下游通路。研究发现，miR-212-5p在NPC组织中表达下调，miR-212-5p低表达与淋巴结转移和低分化相关。miR-212-5p过表达抑制NPC细胞生长并促进其凋亡；miR-212-5p抑制则起相反作用。机制上，miR-212-5p通过抑制METTL3表达来抑制NPC细胞增殖并促进其凋亡。miR-212-5p/METTL3与RNA转运和细胞周期过程相关。总之，miR-212-5p通过靶向METTL3抑制NPC的进展。
