E2F8 Induces Cell Proliferation and Invasion through the Epithelial–Mesenchymal Transition and Notch Signaling Pathways in Ovarian Cancer
Abstract
:1. Introduction
2. Results
2.1. E2F8 Expression Elevated in Ovarian Cancer and Correlated with Poor Prognosis
2.2. E2F8 Expression Elevated in Ovarian Cancer Cell Lines and Correlated with Cell Proliferation
2.3. E2F8 Knockdown Inhibits Invasion and Migration of Ovarian Cancer Cells
2.4. E2F8 Knockdown Blocked Tumor Growth in Xenograft Nude Mouse Model
2.5. Effects of E2F8 Knockdown on EMT and Notch Pathways
3. Disussion
4. Methods
4.1. Patients and Tissue Samples
4.2. Ethics Approval and Consent to Participate
4.3. Tumor Microarray (TMA) Construction and Immunohistochemistry (IHC)
4.4. Cell Lines
4.5. Small Interfering RNA (siRNA) Transfection
4.6. Construction of Lentiviral E2F8-shRNA Vector
4.7. Quantitative Real-Time PCR (qRT-PCR)
4.8. Cell Proliferation Assay
4.9. Clonogenic Survival Assay
4.10. Wound-Healing Migration Assay
4.11. Matrigel Invasion Assay
4.12. Western Blotting Analysis
4.13. In-Vivo Experiment
4.14. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
E2F8 | E2F transcription factor 8 |
EMT | epithelial–mesenchymal transition |
TMA | tumor microarray |
IHC | immunohistochemistry |
KCLB | Korean Cell Line Bank |
ECACC | European Collection of Cell Cultures |
HOSE | human ovarian surface epithelial cells |
OEpiCM | varian epithelial cell medium |
siRNA | small interfering RNA |
siE2F8 | E2F8-targeting siRNA |
siNC | negative control siRNA |
shE2F8.1 and shE2F8.3 | shRNAs targeting E2F8 |
qRT-PCR | quantitative real-time PCR |
CCK-8 | Cell Counting Kit-8 |
RIPA | radioimmunoprecipitation assay |
FIGO | International Federation of Gynecology and Obstetrics |
NRF | National Research Foundation of Korea |
HR | hazard ratio |
CI | confidence interval |
CA | cancer antigen |
NGR | no gross residual; |
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No. of Patients | Progression-Free Survival | ||||
---|---|---|---|---|---|
Univariate Analysis | Multivariate Analysis | ||||
HR (95% CI) | p | HR (95% CI) | p | ||
Age, years (continuous) | 115 | 0.996 (0.975–1.017) | 0.683 | ||
FIGO stage | |||||
I, II | 11 | 1.0 | 1.0 | ||
III, IV | 104 | 5.519 (1.318–23.1.6) | 0.019 | 5.361 (1.293–22.224) | 0.021 |
Tumor grade | |||||
Low | 4 | 1.0 | |||
High | 111 | 3.614 (0.484–26.996) | 0.21 | ||
Preoperative CA 125, U/mL | |||||
≤500 | 51 | 1.0 | |||
>500 | 64 | 1.004 (0.620–1.625) | 0.987 | ||
Residual disease | |||||
NGR | 75 | 1.0 | 1.0 | ||
R < 1 cm | 40 | 17.787 (8.400–37.664) | <0.001 | 19.029 (9.030–40.100) | <0.001 |
E2F8 expression | |||||
Low | 34 | 1.0 | 1.0 | ||
High | 81 | 1.887 (1.073–3.318) | 0.028 | 1.590 (1.010–2.500) | 0.015 |
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Eoh, K.J.; Kim, H.J.; Lee, J.W.; Kim, L.K.; Park, S.-A.; Kim, H.-S.; Kim, Y.T.; Koo, P.J. E2F8 Induces Cell Proliferation and Invasion through the Epithelial–Mesenchymal Transition and Notch Signaling Pathways in Ovarian Cancer. Int. J. Mol. Sci. 2020, 21, 5813. https://doi.org/10.3390/ijms21165813
Eoh KJ, Kim HJ, Lee JW, Kim LK, Park S-A, Kim H-S, Kim YT, Koo PJ. E2F8 Induces Cell Proliferation and Invasion through the Epithelial–Mesenchymal Transition and Notch Signaling Pathways in Ovarian Cancer. International Journal of Molecular Sciences. 2020; 21(16):5813. https://doi.org/10.3390/ijms21165813
Chicago/Turabian StyleEoh, Kyung Jin, Hee Jung Kim, Jong Woo Lee, Lee Kyung Kim, Sun-Ae Park, Hyun-Soo Kim, Young Tae Kim, and Peter J. Koo. 2020. "E2F8 Induces Cell Proliferation and Invasion through the Epithelial–Mesenchymal Transition and Notch Signaling Pathways in Ovarian Cancer" International Journal of Molecular Sciences 21, no. 16: 5813. https://doi.org/10.3390/ijms21165813