Abstract
Background
Ovarian cancer is one of the most common malignant gynecological cancers. Higher expression of AURKA has been found in immortalized human ovarian epithelial cells in previous studies, implying the relationship between AURKA and ovarian cancer pathogenesis.
Aim
We investigated the effect of AURKA on angiogenesis and tumorigenesis of human ovarian cancer cells.
Methods
Firstly, the expression of AURKA in HO8910 and SKOV3 ovarian cancer cell lines was knocked down using a vector expressing a short hairpin small interfering RNA (shRNA). Next, the effect of knockdown of AURKA on cell angiogenesis, proliferation, migration, and invasion was determined by microtubule formation assay, proliferation assay, transwell migration, and invasion assays. In addition, the effect of AURKA knockdown on angiogenesis and tumorigenesis was also determined in a chicken chorioallantoic membrane (CAM) model and in nude mice.
Results
The results of the microtubule formation assay indicated that knockdown of AURKA significantly inhibited ovarian cancer cell-induced angiogenesis of endothelial cells compared to its control (P < 0.001). Knockdown of AURKA also significantly inhibited cell proliferation, migration, and invasion of HO8910 and SKOV3 cells in vitro. Furthermore, the Matrigel plug assay showed that knockdown of AURKA significantly repressed ovarian cancer cell-induced angiogenesis in nude mice (P < 0.05), and the CAMs model also showed that AURKA knockdown significantly attenuated the angiogenesis (P < 0.001) and tumorigenesis (P < 0.001) of HO8910 cells compared to the control. Finally, the tumorigenicity assay in vivo further indicated that AURKA shRNA reduced tumorigenesis in nude mice inoculated with ovarian cancer cells (P < 0.001).
Conclusions
These results suggest the potential role of AURKA in angiogenesis and tumorigenesis of ovarian cancer, which may provide a potential therapeutic target for the disease.
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Conflict of Interest
Zhenqing Feng, Cong Wang, Qin Yan, Minmin Hu, and Di Qin have nothing to disclose.
Funding Statements
This work was supported by grants from National Natural Science Foundation of China (81201703).
Ethical Standards
The animal research has been reviewed and ethically approved by the Institutional Ethics Committee of Nanjing Medical University (Nanjing, China).
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Cong Wang and Qin Yan contributed equally to this work
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Figure S1
Lower expression of AURKA decreased the angiogenesis of endothelial cells induced by HO8910. A Western blot analysis of the expression of AURKA in HO8910 cells transfected with the mixture of siRNA (siRNA) or the negative control (NC). B Matrigel assay analysis of tube formation in EA.hy926 cells cultured with the supernatant from HO8910 cells treated as in A. The representative images of microtubules were taken at 16 h post incubation (original magnification, ×100). C Quantification of results in B. Data represent mean ± SD from three independent experiments (n = 3). ***P < 0.001 for Student’s t test. (PDF 126 kb)
Figure S2
Lower expression of AURKA decreased the angiogenesis of endothelial cells induced by SKOV3. A Western blot analysis of the expression of AURKA in SKOV3 transfected with the mixture of siRNA (siRNA) or the negative control (NC). B Matrigel assay analysis of tube formation in EA.hy926 cells cultured with the supernatant of SKOV3 cells treated as in A. The representative images of microtubules were taken at 16 h post incubation (original magnification, ×100). C Quantification of results in B. Data represent mean ± SD from three independent experiments (n = 3). ***P < 0.001 for Student’s t test. (PDF 122 kb)
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Wang, C., Yan, Q., Hu, M. et al. Effect of AURKA Gene Expression Knockdown on Angiogenesis and Tumorigenesis of Human Ovarian Cancer Cell Lines. Targ Oncol 11, 771–781 (2016). https://doi.org/10.1007/s11523-016-0436-7
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DOI: https://doi.org/10.1007/s11523-016-0436-7