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[Analysis the mechanism of cadmium-induced cytotoxicity in mouse testicular mesenchymal cells with miRNA
Yajing Pian1, Haolong Luo1, Rui Lin1
1Department of Nutrition, School of Public Health, Xuzhou Medical University, Xuzhou 221004, China.
Objective:
To analyze the mechanism of cadmium-induced cytotoxicity in mouse testicular mesenchymal cells(TM3) with transcriptome sequencing and bioinformatics techniques.
Methods:
TM3 cells were selected as the cell model and divided into control group(no cadmium treatment) and cadmium-treated group(20 μmol/L CdCl_2). After 24 hours of administration, cells were harvested to extract total RNA, and then miRNA expression profiles were obtained by sequencing program after RNA quality detection. The fold change(FC) >2, P<0.05 was used as the standard to screen for differentially expressed miRNAs. The quantitative real-time polymerase chain reaction(qRT-PCR) was used to verify the differentially expressed miRNAs. Then, their target genes were predicted by miRanda software to construct miRNA-target gene interaction network, and their target genes were enriched by gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) pathway function.
Results:
A total of 26 differentially expressed miRNAs were identified which may be related to cadmium-induced TM3 cytotoxicity, including 19 up-regulated and 7 down-regulated miRNAs. The result of qRT-PCR were consistent with the miRNA sequencing result. Meanwhile, bioinformatics analysis result showed that the 26 differentially expressed miRNAs predicted 657 target genes. GO enrichment was mainly classified into biological regulation, metabolic process, protein binding and catalytic activity. KEGG pathway analysis showed that target genes were significantly involved in mitogen-activated protein kinase(MAPK) and tumor necrosis factor(TNF) signal pathways closely related to inflammatory response and apoptosis.
Conclusion:
Cadmium can lead to the differential expression of miRNAs in TM3 cells, and its target genes may be involved in Cd-induced TM3 cytotoxicity through signaling pathways such as MAPK and TNF.
Insights
Cadmium exposure alters microRNA (miRNA) expression in mouse testicular cells, impacting biological regulation and signaling pathways. These changes are linked to cadmium-induced cytotoxicity, highlighting potential mechanisms of toxicity.
Area of Science:
- Molecular Biology
- Toxicology
- Genomics
Context:
- Cadmium is a toxic heavy metal with known adverse effects on reproductive health.
- Understanding the molecular mechanisms of cadmium toxicity is crucial for developing protective strategies.
- Testicular mesenchymal cells (TM3) are a relevant model for studying cadmium's impact on testicular function.
Purpose:
- To investigate the role of microRNAs (miRNAs) in cadmium-induced cytotoxicity in mouse testicular mesenchymal cells (TM3).
- To identify differentially expressed miRNAs and their target genes using transcriptome sequencing and bioinformatics.
- To elucidate the signaling pathways involved in cadmium toxicity.
Summary:
- Transcriptome sequencing identified 26 differentially expressed miRNAs (19 up-regulated, 7 down-regulated) in TM3 cells after cadmium exposure.
- Bioinformatics analysis predicted 657 target genes for these miRNAs.
- Gene Ontology and KEGG pathway analyses revealed significant enrichment in biological regulation, metabolic processes, and signaling pathways like MAPK and TNF.
Impact:
- This study reveals that cadmium exposure induces significant miRNA alterations in testicular cells.
- The identified miRNAs and their target genes, particularly those in MAPK and TNF pathways, are implicated in cadmium-induced cytotoxicity.
- Findings provide novel insights into the molecular mechanisms of cadmium's reproductive toxicity.
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