Eugenol inhibits NEAT1 as a ceRNA in pre-cancerous breast lesions
Yusheng Liu1,2, Guijuan Zhang2, Yi Ma3
1Department of Traditional Chinese Medicine, Yangjiang People's Hospital, Yangjiang, Guangdong, 529525, China.
Objective:
Eugenol (EU) from cloves is highly effective against different tumors. The long noncoding ribonucleic acids (lncRNAs), which play a role of competing endogenous RNAs (ceRNAs), suppress microRNAs (miRNAs) involved in post-transcriptional regulatory networks. The present work focused on analyzing how EU affected pre-cancerous breast lesions (PBL).
Methods:
Initially, the gene expression profiles of patients (n = 880) in the National Center for Biotechnology Information (NCBI) database were analyzed. Further, we established a lncRNA-miRNA-mRNA ceRNA network through bioinformatics analysis and investigated mechanistic roles of lncRNAs as ceRNAs and the anti-tumor effect of EU using MCF-10AT cells in vitro as well as PBL model rats in vivo. Besides, Nuclear Paraspeckle Assembly Transcript 1 (NEAT1), miR-383-5p, miR-9-5p, matrix metalloproteinase-9 (MMP-9), and vascular endothelial growth factor-A (VEGF-A) expression was examined through quantitative reverse transcription polymerase chain reaction (RT-qPCR), Western blotting, and immunohistochemical staining analyses.
Results:
There were altogether 1162 mRNAs, 81 miRNAs, and 26 lncRNAs recognized as trend genes in breast cancer (BC) and pre-cancerous BC (pBC), constructing the ceRNA network using 3 lncRNAs, 3 miRNAs, and 38 mRNAs. It was observed that NEAT1, miR-383-5p, miR-9-5p, VEGF-A, and MMP-9 were downregulated in breast tumor cells in accordance with bioinformatics analysis. EU suppressed MCF-10AT cell growth, decreasing the NEAT1, VEGF-A, and MMP-9 levels and increasing miR-383-5p and miR-9-5p expressions in vitro and in vivo.
Conclusion:
In summary, the EU reduced the VEGF-A and MMP-9 expressions via NEAT1-mediated miR-383-5p and miR-9-5p against PBL, indicating that the EU may be a promising external drug to act against PBL.
Insights
Eugenol (EU), a compound from cloves, shows promise in treating pre-cancerous breast lesions (PBL). It works by downregulating NEAT1, VEGF-A, and MMP-9 while upregulating miR-383-5p and miR-9-5p, offering a potential therapeutic strategy.
Area of Science:
- Molecular biology
- Oncology
- Pharmacology
Background:
- Long noncoding RNAs (lncRNAs) function as competing endogenous RNAs (ceRNAs), regulating microRNAs (miRNAs) in post-transcriptional networks.
- Eugenol (EU), derived from cloves, exhibits significant anti-tumor properties.
- Pre-cancerous breast lesions (PBL) represent an important stage in breast cancer development.
Purpose of the Study:
- To investigate the effects of Eugenol (EU) on pre-cancerous breast lesions (PBL).
- To elucidate the role of the lncRNA-miRNA-mRNA ceRNA network in EU's anti-tumor activity.
- To analyze the expression of key molecules including NEAT1, miR-383-5p, miR-9-5p, MMP-9, and VEGF-A in response to EU treatment.
Main Methods:
- Bioinformatic analysis of gene expression profiles from the NCBI database (n=880).
- Construction of a lncRNA-miRNA-mRNA ceRNA network.
- In vitro studies using MCF-10AT cells and in vivo studies using a PBL rat model.
- Quantitative reverse transcription polymerase chain reaction (RT-qPCR), Western blotting, and immunohistochemical staining to assess gene and protein expression.
Main Results:
- A ceRNA network was constructed involving 3 lncRNAs, 3 miRNAs, and 38 mRNAs, identifying 1162 mRNAs, 81 miRNAs, and 26 lncRNAs as trend genes in breast cancer and pre-cancerous breast cancer.
- NEAT1, miR-383-5p, miR-9-5p, VEGF-A, and MMP-9 were found to be downregulated in breast tumor cells.
- EU treatment suppressed MCF-10AT cell growth, decreased NEAT1, VEGF-A, and MMP-9 levels, and increased miR-383-5p and miR-9-5p expression both in vitro and in vivo.
Conclusions:
- Eugenol (EU) effectively suppresses pre-cancerous breast lesions (PBL) by modulating the NEAT1-mediated ceRNA network involving miR-383-5p and miR-9-5p.
- EU reduces VEGF-A and MMP-9 expression through this pathway.
- EU demonstrates potential as a therapeutic agent for PBL.
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