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Ferroptosis Altered microRNAs Expression in HT-1080 Fibrosarcoma Cells Based on Small RNA Sequencing and
Qian Zhang1, Qiwen Wang1, Haoxuan Ding1
1Key Laboratory of Animal Nutrition and Feed of Zhejiang Province, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Abstract:
Iron is an essential trace element in the human body. However, excess iron is harmful and may cause ferroptosis. The expression and role of microRNAs (miRNAs) in ferroptosis remain largely unknown. A model of ferroptosis induced by ferric ammonium citrate in HT-1080 cells was established in this study. The miRNAs expression profiles of the control and iron groups were obtained using small RNA sequencing and verified using qRT-PCR. A total of 1346 known miRNAs and 80 novel miRNAs were identified, including 12 up-regulated differentially expressed miRNAs (DE-miRNAs) and 16 down-regulated DE-miRNAs. SP1 was the most important upstream transcription factor regulating DE-miRNAs. The downstream target genes of DE-miRNAs were predicted based on miRDB, TargetScan, and miRBase databases, and 403 common target genes were screened. GO annotation and KEGG analysis revealed that the target genes were mainly involved in various biological processes and regulatory pathways, especially the MAPK signaling pathway and PI3K-Akt signaling pathway. Afterwards, a target genes network was constructed using STRING and Cytoscape, and the hub genes were compared with the ferroptosis database (FerrDb V2) to discover the hub genes related to ferroptosis. EGFR, GSK3B, PARP1, VCP, and SNCA were screened out. Furthermore, a DE-miRNAs-target genes network was constructed to explore key DE-miRNAs. hsa-miR-200c-3p, hsa-miR-26b-5p, and hsa-miR-7-5p were filtered out. Comprehensive bioinformatics analysis of miRNAs and its upstream and downstream regulation in ferroptosis in HT-1080 cells using small RNA sequencing is helpful for understanding the role of miRNAs in iron overload-related diseases and ferroptosis-targeted therapy for cancer.
Insights
This study reveals microRNAs (miRNAs) involved in iron overload-induced ferroptosis. Key miRNAs and target genes like EGFR were identified, offering insights into ferroptosis-targeted cancer therapy.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Iron is essential but toxic in excess, potentially causing ferroptosis.
- The role and expression of microRNAs (miRNAs) in ferroptosis are largely unknown.
- Understanding miRNA involvement is crucial for iron overload diseases and cancer therapy.
Purpose of the Study:
- To investigate the expression profiles and regulatory roles of miRNAs in ferroptosis.
- To identify key miRNAs, their upstream regulators, and downstream targets in iron-induced ferroptosis.
- To explore potential therapeutic targets for ferroptosis-related conditions.
Main Methods:
- Ferroptosis model induced by ferric ammonium citrate in HT-1080 cells.
- Small RNA sequencing for miRNA expression profiling, validated by qRT-PCR.
- Bioinformatic analyses including target gene prediction, pathway analysis (GO, KEGG), and network construction (STRING, Cytoscape).
Main Results:
- Identified 1346 known and 80 novel miRNAs, with 12 up-regulated and 16 down-regulated DE-miRNAs.
- SP1 identified as a key upstream regulator; 403 common target genes predicted.
- Key ferroptosis-related hub genes (EGFR, GSK3B, PARP1, VCP, SNCA) and DE-miRNAs (hsa-miR-200c-3p, hsa-miR-26b-5p, hsa-miR-7-5p) were screened.
Conclusions:
- Comprehensive bioinformatics analysis elucidated miRNA regulation in ferroptosis.
- Identified potential biomarkers and therapeutic targets for iron overload and ferroptosis-related cancers.
- Findings contribute to understanding miRNA roles in iron metabolism and disease pathogenesis.
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