A Novel Estrogen Receptor β Agonist Diminishes Ovarian Cancer Stem Cells via Suppressing the Epithelial-to-Mesenchymal Transition
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Cell Culture and Reagents
2.2. Establishment of Organoid Culture from Freshly Removed Human Ovarian Tumor
2.3. Cell Sensitivity Assay
2.4. RNA Isolation and qPCR Analysis
2.5. Immunoblotting
2.6. Immunofluorescence
2.7. Transwell Migration and Invasion Assays
2.8. ALDH Analysis
2.9. CD44/CD117 Analysis
2.10. Small Interference RNA (siRNA) Transfection
2.11. Sphere Forming Assay
2.12. Animal Study
2.13. Statistical Analysis
3. Results
3.1. ERβ Agonist OSU-ERb-12 Is Able to Suppress Ovarian Cancer Cell Proliferation
3.2. ERβ Agonist OSU-ERb-12 Reduces the CSC Population in Ovarian Cancer Cells
3.3. ERβ Agonist OSU-ERb-12 Suppresses the Dedifferentiation of Ovarian Cancer Cells
3.4. ERβ Agonist OSU-ERb-12 Inhibits EMT in Ovarian Cancer Cells
3.5. ERβ Agonist OSU-ERb-12 Limits the CSC Population by Blocking the Function of ERα
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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Genes | Forward | Reverse |
---|---|---|
Nanog | 5′-GTCCCAAAGGCAAACAACCC-3′ | 5′-TTGACCGGGACCTTGTCTTC-3′ |
Sox2 | 5′-TCAGGAGTTGTCAAGGCAGAG-3′ | 5′-GGCAGCAAACTACTTTCCCC-3′ |
Oct4 | 5′-TCGCAAGCCCTCATTTCACC-3′ | 5′-CGAGAAGGCGAAATCCGAAG-3′ |
ESR1 | 5′-GCTACGAAGTGGAATGATGAAAG-3′ | 5′-TCTGGCGCTTGTGTTTCAAC-3′ |
ESR2 | 5′-ACTTGCTGAACGCCGTGACC-3′ | 5′-CAGATGTTCCATGCCCTTGTT-3′ |
GAPDH | 5′-GAAGGTGAAGGTCGGAGT-3′ | 5′-GAAGATGGTGATGGGATTTC-3′ |
CCND1 | 5′-GGGTTGTGCTACAGATGATAGAG-3′ | 5′-GAGGTGACTTCAGCCATGAATA-3′ |
NRIP1 | 5′-CTCCAAGAATGGTCTGCTAAGT-3′ | 5′-GGTTAAGCAAGGACCCATACA-3′ |
Antibody | Catalog Number | Company | Dilution Factor |
---|---|---|---|
Anti-ERα | #8644S | Cell Signaling Technology | 1:500 |
Anti-ERß | N/A | Ohio State University | 1:500 |
Anti-cPARP | #9546S | Cell Signaling Technology | 1:1000 |
Anti-GAPDH | #Sc-365062 | Santa Cruz Biotechnology | 1:2000 |
Anti-Tubulin | #2144S | Cell Signaling Technology | 1:2000 |
Anti-E-Cadherin | #610077 | BD Transduction Laboratories | 1:1000 |
Anti-Snail | #3879S | Cell Signaling Technology | 1:1000 |
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Banerjee, A.; Cai, S.; Xie, G.; Li, N.; Bai, X.; Lavudi, K.; Wang, K.; Zhang, X.; Zhang, J.; Patnaik, S.; et al. A Novel Estrogen Receptor β Agonist Diminishes Ovarian Cancer Stem Cells via Suppressing the Epithelial-to-Mesenchymal Transition. Cancers 2022, 14, 2311. https://doi.org/10.3390/cancers14092311
Banerjee A, Cai S, Xie G, Li N, Bai X, Lavudi K, Wang K, Zhang X, Zhang J, Patnaik S, et al. A Novel Estrogen Receptor β Agonist Diminishes Ovarian Cancer Stem Cells via Suppressing the Epithelial-to-Mesenchymal Transition. Cancers. 2022; 14(9):2311. https://doi.org/10.3390/cancers14092311
Chicago/Turabian StyleBanerjee, Ananya, Shurui Cai, Guozhen Xie, Na Li, Xuetao Bai, Kousalya Lavudi, Kevin Wang, Xiaoli Zhang, Junran Zhang, Srinivas Patnaik, and et al. 2022. "A Novel Estrogen Receptor β Agonist Diminishes Ovarian Cancer Stem Cells via Suppressing the Epithelial-to-Mesenchymal Transition" Cancers 14, no. 9: 2311. https://doi.org/10.3390/cancers14092311