1α,25(OH)2D3 Suppresses the Migration of Ovarian Cancer SKOV-3 Cells through the Inhibition of Epithelial–Mesenchymal Transition
Abstract
:1. Introduction
2. Results
2.1. 1α,25(OH)2D3 Inhibits the Migration of Human Ovarian Cancer SKOV-3 Cells
2.2. Establishment of TGF-β1-Induced EMT in SKOV-3 Cells
2.3. 1α,25 (OH)2D3 Inhibited the Migration and Invasion of SKOV-3 Cells during TGF-β1-Induced EMT
2.4. 1α,25(OH)2D3 Regulates the Expression of EMT Markers in TGF-β1-Treated SKOV-3 Cells
3. Discussion
4. Experimental Section
4.1. Cell Culture and Treatment
4.2. Wound Healing Assay
4.3. Live Cell Imaging System
4.4. Invasion Assay
4.5. Immunofluoresecence Staining
4.6. Western Blot
4.7. Statistical Analysis
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
EMT | Epithelial-mesenchymal transition |
1α,25(OH)2D3 | 1α,25-dihydroxyvitamin D |
VDR | vitamin D receptor |
25(OH)D | 25-dihydroxyvitamin D |
References
- Siegel, R.; Ward, E.; Brawley, O.; Jemal, A. Cancer statistics, 2011: The impact of eliminating socioeconomic and racial disparities on premature cancer deaths. CA Cancer J. Clin. 2011, 61, 212–236. [Google Scholar] [CrossRef] [PubMed]
- Lamouille, S.; Xu, J.; Derynck, R. Molecular mechanisms of epithelial-mesenchymal transition. Nat. Rev. Mol. Cell Biol. 2014, 15, 178–196. [Google Scholar] [CrossRef] [PubMed]
- Ahmed, N.; Thompson, E.W.; Quinn, M.A. Epithelial-mesenchymal interconversions in normal ovarian surface epithelium and ovarian carcinomas: An exception to the norm. J. Cell. Physiol. 2007, 213, 581–588. [Google Scholar] [CrossRef] [PubMed]
- Yan, H.; Sun, Y. Evaluation of the mechanism of epithelial-mesenchymal transition in human ovarian cancer stem cells transfected with a WW domain-containing oxidoreductase gene. Oncol. Lett. 2014, 8, 426–430. [Google Scholar] [CrossRef] [PubMed]
- Abell, A.N.; Johnson, G.L. Implications of mesenchymal cells in cancer stem cell populations: Relevance to emt. Curr. Pathobiol. Rep. 2014, 2, 21–26. [Google Scholar] [CrossRef] [PubMed]
- Li, P.; Li, C.; Zhao, X.; Zhang, X.; Nicosia, S.V.; Bai, W. P27(Kip1) stabilization and G(1) arrest by 1,25-dihydroxyvitamin D(3) in ovarian cancer cells mediated through down-regulation of cyclin e/cyclin-dependent kinase 2 and Skp1-Cullin-F-box protein/Skp2 ubiquitin ligase. J. Biol. Chem. 2004, 279, 25260–25267. [Google Scholar] [CrossRef] [PubMed]
- Deeb, K.K.; Trump, D.L.; Johnson, C.S. Vitamin D signalling pathways in cancer: Potential for anticancer therapeutics. Nat. Rev. Cancer 2007, 7, 684–700. [Google Scholar] [CrossRef] [PubMed]
- Feldman, D.; Krishnan, A.V.; Swami, S.; Giovannucci, E.; Feldman, B.J. The role of vitamin D in reducing cancer risk and progression. Nat. Rev. Cancer 2014, 14, 342–357. [Google Scholar] [CrossRef] [PubMed]
- Ahonen, M.H.; Zhuang, Y.H.; Aine, R.; Ylikomi, T.; Tuohimaa, P. Androgen receptor and vitamin D receptor in human ovarian cancer: Growth stimulation and inhibition by ligands. Int. J. Cancer 2000, 86, 40–46. [Google Scholar] [CrossRef]
- Saunders, D.E.; Christensen, C.; Wappler, N.L.; Schultz, J.F.; Lawrence, W.D.; Malviya, V.K.; Malone, J.M.; Deppe, G. Inhibition of c-myc in breast and ovarian carcinoma cells by 1,25-dihydroxyvitamin D3, retinoic acid and dexamethasone. Anticancer Drugs 1993, 4, 201–208. [Google Scholar] [CrossRef] [PubMed]
- Jiang, F.; Bao, J.; Li, P.; Nicosia, S.V.; Bai, W. Induction of ovarian cancer cell apoptosis by 1,25-dihydroxyvitamin D3 through the down-regulation of telomerase. J. Biol. Chem. 2004, 279, 53213–53221. [Google Scholar] [CrossRef] [PubMed]
- Zhang, Z.; Zhang, H.; Hu, Z.; Wang, P.; Wan, J.; Li, B. Synergy of 1,25-dihydroxyvitamin D3 and carboplatin in growth suppression of SKOV-3 cells. Oncol. Lett. 2014, 8, 1348–1354. [Google Scholar] [CrossRef] [PubMed]
- Lange, T.S.; Stuckey, A.R.; Robison, K.; Kim, K.K.; Singh, R.K.; Raker, C.A.; Brard, L. Effect of a vitamin D(3) derivative (B3CD) with postulated anti-cancer activity in an ovarian cancer animal model. Investig. New Drugs 2010, 28, 543–553. [Google Scholar] [CrossRef] [PubMed]
- Kawar, N.; Maclaughlan, S.; Horan, T.C.; Uzun, A.; Lange, T.S.; Kim, K.K.; Hopson, R.; Singh, A.P.; Sidhu, P.S.; Glass, K.A.; et al. Pt19c, another nonhypercalcemic vitamin D2 derivative, demonstrates antitumor efficacy in epithelial ovarian and endometrial cancer models. Genes Cancer 2013, 4, 524–534. [Google Scholar] [CrossRef] [PubMed]
- Chen, S.; Zhu, J.; Zuo, S.; Ma, J.; Zhang, J.; Chen, G.; Wang, X.; Pan, Y.; Liu, Y.; Wang, P. 1,25(OH)2D3 attenuates Tgf-beta1/beta2-induced increased migration and invasion via inhibiting epithelial-mesenchymal transition in colon cancer cells. Biochem. Biophys. Res. Commun. 2015, 468, 130–135. [Google Scholar] [CrossRef] [PubMed]
- Davidson, B.; Trope, C.G.; Reich, R. Epithelial-mesenchymal transition in ovarian carcinoma. Front. Oncol. 2012, 2, 33. [Google Scholar] [CrossRef] [PubMed]
- Chiang, K.C.; Chen, S.C.; Yeh, C.N.; Pang, J.H.; Shen, S.C.; Hsu, J.T.; Liu, Y.Y.; Chen, L.W.; Kuo, S.F.; Takano, M.; et al. MART-10, a less calcemic vitamin D analog, is more potent than 1alpha,25-dihydroxyvitamin D3 in inhibiting the metastatic potential of MCF-7 breast cancer cells in vitro. J. Steroid Biochem. Mol. Biol. 2014, 139, 54–60. [Google Scholar] [CrossRef] [PubMed]
- Chiang, K.C.; Yeh, C.N.; Hsu, J.T.; Jan, Y.Y.; Chen, L.W.; Kuo, S.F.; Takano, M.; Kittaka, A.; Chen, T.C.; Chen, W.T.; et al. The vitamin D analog, MART-10, represses metastasis potential via downregulation of epithelial-mesenchymal transition in pancreatic cancer cells. Cancer Lett. 2014, 354, 235–244. [Google Scholar] [CrossRef] [PubMed]
- Upadhyay, S.K.; Verone, A.; Shoemaker, S.; Qin, M.; Liu, S.; Campbell, M.; Hershberger, P.A. 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) signaling capacity and the epithelial-mesenchymal transition in non-small cell lung cancer (NSCLC): Implications for use of 1,25(OH)2D3 in NSCLC treatment. Cancers 2013, 5, 1504–1521. [Google Scholar] [CrossRef] [PubMed]
- Wong, A.S.; Auersperg, N. Normal ovarian surface epithelium. Cancer Treat. Res. 2002, 107, 161–183. [Google Scholar] [PubMed]
- Sundfeldt, K.; Piontkewitz, Y.; Ivarsson, K.; Nilsson, O.; Hellberg, P.; Brannstrom, M.; Janson, P.O.; Enerback, S.; Hedin, L. E-cadherin expression in human epithelial ovarian cancer and normal ovary. Int. J. Cancer 1997, 74, 275–280. [Google Scholar] [CrossRef]
- Zavadil, J.; Bottinger, E.P. Tgf-beta and epithelial-to-mesenchymal transitions. Oncogene 2005, 24, 5764–5774. [Google Scholar] [CrossRef] [PubMed]
- Pang, M.F.; Georgoudaki, A.M.; Lambut, L.; Johansson, J.; Tabor, V.; Hagikura, K.; Jin, Y.; Jansson, M.; Alexander, J.S.; Nelson, C.M.; et al. TGF-beta1-induced EMT promotes targeted migration of breast cancer cells through the lymphatic system by the activation of CCR7/CCL21-mediated chemotaxis. Oncogene 2016, 35, 748–760. [Google Scholar] [CrossRef] [PubMed]
- Vincent, T.; Neve, E.P.; Johnson, J.R.; Kukalev, A.; Rojo, F.; Albanell, J.; Pietras, K.; Virtanen, I.; Philipson, L.; Leopold, P.L.; et al. A SNAIL1-SMAD3/4 transcriptional repressor complex promotes tgf-beta mediated epithelial-mesenchymal transition. Nat. Cell Biol. 2009, 11, 943–950. [Google Scholar] [CrossRef] [PubMed]
- Goswami, M.T.; Reka, A.K.; Kurapati, H.; Kaza, V.; Chen, J.; Standiford, T.J.; Keshamouni, V.G. Regulation of complement-dependent cytotoxicity by TGF-beta-induced epithelial-mesenchymal transition. Oncogene 2016, 35, 1888–1898. [Google Scholar] [CrossRef] [PubMed]
- Liu, R.Y.; Zeng, Y.; Lei, Z.; Wang, L.; Yang, H.; Liu, Z.; Zhao, J.; Zhang, H.T. JAK/STAT3 signaling is required for TGF-beta-induced epithelial-mesenchymal transition in lung cancer cells. Int. J. Oncol. 2014, 44, 1643–1651. [Google Scholar] [PubMed]
- Leyssens, C.; Verlinden, L.; Verstuyf, A. Antineoplastic effects of 1,25(OH)2D3 and its analogs in breast, prostate and colorectal cancer. Endocr. Relat. Cancer 2013, 20, R31–R47. [Google Scholar] [CrossRef] [PubMed]
- Villena-Heinsen, C.; Meyberg, R.; Axt-Fliedner, R.; Reitnauer, K.; Reichrath, J.; Friedrich, M. Immunohistochemical analysis of 1,25-dihydroxyvitamin-D3-receptors, estrogen and progesterone receptors and Ki-67 in ovarian carcinoma. Anticancer Res. 2002, 22, 2261–2267. [Google Scholar] [PubMed]
- Zheng, W.; Danforth, K.N.; Tworoger, S.S.; Goodman, M.T.; Arslan, A.A.; Patel, A.V.; McCullough, M.L.; Weinstein, S.J.; Kolonel, L.N.; Purdue, M.P.; et al. Circulating 25-hydroxyvitamin D and risk of epithelial ovarian cancer: Cohort consortium vitamin D pooling project of rarer cancers. Am. J. Epidemiol. 2010, 172, 70–80. [Google Scholar] [CrossRef] [PubMed]
- Yin, L.; Grandi, N.; Raum, E.; Haug, U.; Arndt, V.; Brenner, H. Meta-analysis: Circulating vitamin D and ovarian cancer risk. Gynecol. Oncol. 2011, 121, 369–375. [Google Scholar] [CrossRef] [PubMed]
- Jiang, F.; Li, P.; Fornace, A.J., Jr.; Nicosia, S.V.; Bai, W. G2/M arrest by 1,25-dihydroxyvitamin D3 in ovarian cancer cells mediated through the induction of GADD45 via an exonic enhancer. J. Biol. Chem. 2003, 278, 48030–48040. [Google Scholar] [CrossRef] [PubMed]
- Chiang, K.C.; Yeh, T.S.; Chen, S.C.; Pang, J.H.; Yeh, C.N.; Hsu, J.T.; Chen, L.W.; Kuo, S.F.; Takano, M.; Kittaka, A.; et al. The vitamin D analog, MART-10, attenuates triple negative breast cancer cells metastatic potential. Int. J. Mol. Sci. 2016, 17, 606. [Google Scholar] [CrossRef] [PubMed]
- Lee, J.M.; Dedhar, S.; Kalluri, R.; Thompson, E.W. The epithelial-mesenchymal transition: New insights in signaling, development, and disease. J. Cell Biol. 2006, 172, 973–981. [Google Scholar] [CrossRef] [PubMed]
- Egan, J.B.; Thompson, P.A.; Vitanov, M.V.; Bartik, L.; Jacobs, E.T.; Haussler, M.R.; Gerner, E.W.; Jurutka, P.W. Vitamin D receptor ligands, adenomatous polyposis coli, and the vitamin D receptor foki polymorphism collectively modulate beta-catenin activity in colon cancer cells. Mol. Carcinog. 2010, 49, 337–352. [Google Scholar] [PubMed]
- Larriba, M.J.; Munoz, A. Snail versus vitamin D receptor expression in colon cancer: Therapeutics implications. Br. J. Cancer 2005, 92, 985–989. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Pena, C.; Garcia, J.M.; Garcia, V.; Silva, J.; Dominguez, G.; Rodriguez, R.; Maximiano, C.; Garcia de Herreros, A.; Munoz, A.; Bonilla, F. The expression levels of the transcriptional regulators p300 and CtBP modulate the correlations between SNAIL, ZEB1, E-cadherin and vitamin D receptor in human colon carcinomas. Int. J. Cancer 2006, 119, 2098–2104. [Google Scholar] [CrossRef] [PubMed]
© 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Hou, Y.-F.; Gao, S.-H.; Wang, P.; Zhang, H.-M.; Liu, L.-Z.; Ye, M.-X.; Zhou, G.-M.; Zhang, Z.-L.; Li, B.-Y. 1α,25(OH)2D3 Suppresses the Migration of Ovarian Cancer SKOV-3 Cells through the Inhibition of Epithelial–Mesenchymal Transition. Int. J. Mol. Sci. 2016, 17, 1285. https://doi.org/10.3390/ijms17081285
Hou Y-F, Gao S-H, Wang P, Zhang H-M, Liu L-Z, Ye M-X, Zhou G-M, Zhang Z-L, Li B-Y. 1α,25(OH)2D3 Suppresses the Migration of Ovarian Cancer SKOV-3 Cells through the Inhibition of Epithelial–Mesenchymal Transition. International Journal of Molecular Sciences. 2016; 17(8):1285. https://doi.org/10.3390/ijms17081285
Chicago/Turabian StyleHou, Yong-Feng, Si-Hai Gao, Ping Wang, He-Mei Zhang, Li-Zhi Liu, Meng-Xuan Ye, Guang-Ming Zhou, Zeng-Li Zhang, and Bing-Yan Li. 2016. "1α,25(OH)2D3 Suppresses the Migration of Ovarian Cancer SKOV-3 Cells through the Inhibition of Epithelial–Mesenchymal Transition" International Journal of Molecular Sciences 17, no. 8: 1285. https://doi.org/10.3390/ijms17081285