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In Vivo Inhibition of MicroRNA to Decrease Tumor Growth in Mice
Published on: August 23, 2019
miR-212-5p inhibits nasopharyngeal carcinoma progression by targeting METTL3
1Department of Otorhinolaryngology Head and Neck Surgery, Wuhan Fourth Hospital, Wuhan 430033, Hubei, China.
Abstract:
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.
Insights
MicroRNA-212-5p (miR-212-5p) is downregulated in nasopharyngeal carcinoma (NPC). Restoring miR-212-5p inhibits NPC progression by targeting METTL3, suppressing proliferation and promoting apoptosis.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Nasopharyngeal carcinoma (NPC) is a significant health concern with complex molecular underpinnings.
- MicroRNAs (miRNAs) play crucial roles in cancer development and progression, making them potential therapeutic targets.
- Understanding the specific roles of miRNAs like miR-212-5p in NPC is essential for developing novel treatment strategies.
Purpose of the Study:
- To investigate the functional role of microRNA-212-5p (miR-212-5p) in the proliferation and apoptosis of nasopharyngeal carcinoma (NPC) cells.
- To elucidate the molecular mechanism by which miR-212-5p influences NPC progression, focusing on its potential target genes.
- To explore the association between miR-212-5p expression and clinicopathological features of NPC patients.
Main Methods:
- Analysis of microarray datasets (dbDEMC) to identify differentially expressed miRNAs in NPC.
- Quantitative real-time PCR and Western blot to determine miR-212-5p and METTL3 expression levels.
- In vitro assays (CCK-8, EdU, flow cytometry) to assess cell proliferation and apoptosis.
- Bioinformatics analysis (KEGG, GO, GSEA) and dual-luciferase reporter assay to identify and validate miR-212-5p targets and pathways.
Main Results:
- miR-212-5p was found to be significantly downregulated in NPC tissues and associated with lymph node metastasis and poor differentiation.
- Overexpression of miR-212-5p inhibited NPC cell proliferation and induced apoptosis, while inhibition of miR-212-5p had opposite effects.
- miR-212-5p directly targets methyltransferase like 3 (METTL3), suppressing its expression and thereby inhibiting NPC cell proliferation and promoting apoptosis.
Conclusions:
- miR-212-5p acts as a tumor suppressor in nasopharyngeal carcinoma.
- The miR-212-5p/METTL3 axis plays a critical role in regulating NPC cell proliferation, apoptosis, RNA transport, and cell cycle.
- Restoring miR-212-5p expression represents a potential therapeutic strategy for inhibiting NPC progression.
