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The Analysis of Differentially Expressed circRNAs Under the Antiproliferative Effect From 5-Fluorouracil on
AiJun Huang1, LiPing Chen2, YiMing Wang1
1Department of Orthopaedic Surgery, The Eighth Affiliated Hospital, Sun Yat-Sen University, Shenzhen, People's Republic of China.
Background:
5-fluorouracil (5-FU) is a widely used drug for cancer treatment, but its effect and underlying mechanisms on osteosarcoma (OS) cells remain unclear.
Methods:
U2OS and MG63 cells were treated with 0, 50, 100, and 500 μM 5-FU. MTS and flow cytometry were used to examine the effect of 5-FU on cell viability and apoptosis, respectively. Circular RNA (circRNA) expression was detected using RNA sequencing and quantitative real-time PCR (qPCR). Differentially expressed circRNAs were further subjected to the Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene ontology (GO) analysis to predict their functions. A circRNA-miRNA-mRNA interaction network was generated to analyze the regulatory networks of 5-FU-induced differentially expressed circRNAs. Western blotting (WB) was used to verify the protein in the downstream of circRNAs.
Results:
5-FU inhibited the cell viability of the MG63 cells in a concentration-dependent manner. The most significant effect was observed in the cells treated with 500 μM 5-FU. Apoptosis was also increased in the MG63 cells after 500 μM 5-FU treatment for 3 days. RNA sequencing results showed that 183 differentially expressed circRNAs (172 upregulated and 11 downregulated) in 5-FU-treated cells. KEGG and GO analysis showed that the differentially expressed circRNAs were primarily enriched in proliferation-, apoptosis-, and metabolism-related functions. qPCR was used to verify the most upregulated and downregulated circRNAs. The circRNA-miRNA-mRNA interaction network showed that these 8 circRNAs had a sizable regulatory network that links a series of genes involved in tumor suppression.
Conclusion:
5-FU treatment resulted in the differentially expressed circRNAs that were proliferation- and apoptosis-associated and were involved in the 5-FU-induced inhibition of tumor proliferation in OS cells.
Insights
5-fluorouracil (5-FU) inhibits osteosarcoma cell viability and increases apoptosis. This study identifies differentially expressed circular RNAs (circRNAs) regulated by 5-FU, suggesting their role in tumor suppression.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- 5-fluorouracil (5-FU) is a standard chemotherapy agent.
- Its precise mechanisms in osteosarcoma (OS) remain incompletely understood.
Purpose of the Study:
- To investigate the effects of 5-FU on osteosarcoma cell viability and apoptosis.
- To identify and analyze circular RNAs (circRNAs) differentially expressed in response to 5-FU treatment in OS cells.
Main Methods:
- Osteosarcoma cell lines (U2OS, MG63) were treated with varying concentrations of 5-FU.
- Cell viability and apoptosis were assessed using MTS assays and flow cytometry.
- Circular RNA expression was profiled using RNA sequencing and validated by qPCR.
- Bioinformatic analyses (KEGG, GO) and network construction were performed to predict circRNA functions and interactions.
Main Results:
- 5-FU demonstrated a dose-dependent inhibition of MG63 cell viability and induced apoptosis.
- RNA sequencing identified 183 differentially expressed circRNAs (172 upregulated, 11 downregulated) in 5-FU-treated cells.
- Enriched functions for these circRNAs included proliferation, apoptosis, and metabolism, with a constructed network highlighting tumor-suppressive genes.
Conclusions:
- 5-FU treatment alters circRNA expression profiles in osteosarcoma cells.
- These differentially expressed circRNAs are associated with proliferation and apoptosis, mediating 5-FU's anti-tumor effects in OS.
