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Salinomycin Modulates the Expression of mRNAs and miRNAs Related to Stemness in Endometrial Cancer
Karol Talkowski1, Kamil Kiełbasiński2, Wojciech Peszek3
1Department of Psychiatry, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland.
Background:
Salinomycin, an ionophore antibiotic, has a strong anti-cancer effect, inducing the apoptosis of cancer cells and cancer stem cells.
Objective:
The aim of the study was to assess the influence of salinomycin on the expression profile of genes related to stemness and miRNA regulating their expression in endometrial cancer cells.
Methods:
Endometrial cancer cells of cell line Ishikawa were exposed to salinomycin at concentrations in the range of 0.1-100 μM, with the aim of determining its pro-apoptotic potential and the concentration which would cause the death of 50% of the cells (Sulforhodamine B test). In the following stages, the cells were incubated with the drug at a concentration of 1μM for 12,24 and 48 hour periods and compared to the control. Determining the changes in the expression of the genes related to stemness and regulating their miRNA was done using the microarray technique and RTqPCR. ELISA assay was performed in order to determine the level of TGFβ2, COL14A1, CDH2, WNT5A in cell culture under salinomycin treatment in comparison to the control.
Results:
Salinomycin caused the apoptosis of cells. For the concentration of 0.1 μM, a decrease in the population of living cells by 11.9% was determined. For 1 μM, it was 49.8%, for 10 μM -69.4%, and for a concentration of 100 μM - 87.9%. The most noticeable changes in the expression caused by the addition of salinomycin into the culture were noted for mRNA: TGFβ2; WNT5A (up-regulated); COL14A1; CDH2 (down-regulated), as well as miRNA: hsa-miR-411 (up-regulated); hsa-miR-200a; hsa-miR-33a; hsa-miR-199a; hsa-miR-371-5p; hsa-miR-374; hsa-miR-374b (down-regulated).
Conclusion:
It was confirmed that salinomycin has an influence on the stemness process. The most noticeable changes in the expression were noted for mRNA: TGFβ2; COL14A1; CDH2; WNT5A, as well as for miRNA: hsa-miR-200a; hsa-miR-33a; hsa-miR-199a; hsa-miR-371-5p; hsa-miR-411; hsa-miR- 374a; hsa-miR-374b.
Insights
Salinomycin, an ionophore antibiotic, effectively induces apoptosis in endometrial cancer cells and alters the expression of stemness-related genes and microRNAs. This study highlights its potential in cancer therapy by modulating key cellular pathways.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Salinomycin, an ionophore antibiotic, exhibits potent anti-cancer properties by inducing apoptosis in cancer cells and cancer stem cells.
- Endometrial cancer is a significant health concern, necessitating novel therapeutic strategies targeting cancer stemness.
Purpose of the Study:
- To investigate the effect of salinomycin on gene expression profiles related to stemness in endometrial cancer cells.
- To analyze the impact of salinomycin on microRNAs (miRNAs) that regulate stemness-associated genes.
Main Methods:
- Ishikawa endometrial cancer cells were treated with varying concentrations of salinomycin (0.1-100 μM).
- Pro-apoptotic potential was assessed using the Sulforhodamine B assay.
- Gene and miRNA expression changes were analyzed using microarray and RTqPCR techniques.
- Protein levels of TGFβ2, COL14A1, CDH2, and WNT5A were quantified via ELISA.
Main Results:
- Salinomycin demonstrated dose-dependent apoptosis induction, with significant cell death observed at concentrations of 1 μM and higher.
- Upregulation of TGFβ2 and WNT5A mRNA and downregulation of COL14A1 and CDH2 mRNA were observed.
- Specific miRNAs, including hsa-miR-411, were upregulated, while others like hsa-miR-200a, hsa-miR-33a, and hsa-miR-199a were downregulated.
Conclusions:
- Salinomycin influences the stemness processes in endometrial cancer cells.
- The study identified key mRNA and miRNA expression changes associated with salinomycin treatment, providing insights into its anti-cancer mechanisms.
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