Dual inhibition of FOXM1 and its compensatory signaling pathway decreased the survival of ovarian cancer cells

Lee,Dae Woo; Lee,Weonsun; Kwon,Miyeon; Lee,Hae Nam

doi:10.3892/or.2020.7845

Dual inhibition of FOXM1 and its compensatory signaling pathway decreased the survival of ovarian cancer cells

Authors:
- Dae Woo Lee
- Weonsun Lee
- Miyeon Kwon
- Hae Nam Lee
View Affiliations

Affiliations: Department of Obstetrics and Gynecology, Bucheon St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul 14647, Republic of Korea, Clinical Medicine Research Institute, Bucheon St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul 14647, Republic of Korea
Published online on: November 11, 2020 https://doi.org/10.3892/or.2020.7845
Pages: 390-400

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The present study aimed to analyze the compensatory signaling pathways induced by forkhead domain inhibitor‑6 (FDI‑6), which is a forkhead box protein M1 (FOXM1) inhibitor, in ovarian cancer cells and evaluate the effectiveness of simultaneous inhibition of FOXM1 and the compensatory signaling pathway in decreasing the survival of ovarian cancer cells. The present study identified the proteins involved in the compensatory mechanism activated by FDI‑6 in HeyA8 ovarian cancer cells using western blot analysis and a reverse‑phase protein array. In addition, a cell viability assay was performed to determine the effects of FDI‑6 and the compensatory signaling pathway on cancer cell viability. All experiments were performed in three‑dimensional cell cultures. The present study observed that FDI‑6 stimulated the upregulation of N‑Ras, phosphoprotein kinase Cδ (p‑PKCδ) (S664) and HER3 in HeyA8 cells. Tipifarnib as an N‑Ras inhibitor, rottlerin as a p‑PKCδ (S664) inhibitor and sapitinib as a HER3 inhibitor were selected. The combination of FDI‑6 with tipifarnib attenuated the upregulation of N‑Ras induced by FDI‑6 and the combination of FDI‑6 with sapitinib also attenuated HER3 downstream signaling pathway in HeyA8 cells, as shown by on western blot analysis. Rottlerin downregulated p‑PKCδ (S664) by inhibiting the activity of a Src‑related tyrosine kinase that transfers a phosphate group to PKCδ. Compared with FDI‑6 alone, the addition of tipifarnib or rottlerin to FDI‑6 was significantly more effective in reducing the growth of HeyA8 cells. However, the combination of FDI‑6 and sapitinib did not induce a significant decrease in survival of HeyA8 cells. In conclusion, the addition of tipifarnib or rottlerin to inhibit N‑Ras or p‑PKCδ (S664), respectively, inhibited the compensatory signaling pathway response induced by FDI‑6 in HeyA8 cells. These inhibitors increased the efficacy of FDI‑6, which inhibits FOXM1, in reducing ovarian cancer cell viability.

View Figures

View References

1	Lheureux S, Gourley C, Vergote I and Oza AM: Epithelial ovarian cancer. Lancet. 393:1240–1253. 2019. View Article : Google Scholar
2	Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D and Bray F: Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 136:E359–E386. 2015. View Article : Google Scholar
3	Berek JS: Ovarian, fallopian tube and peritoneal cancer. Berek and Novak's Gynecology. 15th edition. Berek JS, Longacre TA and Friedlander M: Wolters Kluwer/Lippincott Williams & Wilkins; Philadelphia, PA: pp. 1350–1427. 2011
4	Cancer Genome Atlas Research Network. Integrated genomic analyses of ovarian carcinoma. Nature. 474:609–615. 2011. View Article : Google Scholar
5	Choi JI, Park SH, Lee HJ, Lee DW and Lee HN: Inhibition of phospho-S6 kinase, a protein involved in the compensatory adaptive response, increases the efficacy of paclitaxel in reducing the viability of matrix-attached ovarian cancer cells. PLoS One. 11:e01550522016. View Article : Google Scholar
6	Gormally MV, Dexheimer TS, Marsico G, Sanders DA, Lowe C, Matak-Vinković D, Michael S, Jadhav A, Rai G, Maloney DJ, et al: Suppression of the FOXM1 transcriptional programme via novel small molecule inhibition. Nat Commun. 5:51652014. View Article : Google Scholar
7	Koo CY, Muir KW and Lam EWF: FOXM1: From cancer initiation to progression and treatment. Biochim Biophys Acta. 1819:28–37. 2012. View Article : Google Scholar
8	Chan DW, Hui WW, Cai PC, Liu MX, Yung MM, Mak CS, Leung TH, Chan KK and Ngan HY: Targeting GRB7/ERK/FOXM1 signaling pathway impairs aggressiveness of ovarian cancer cells. PLoS One. 7:e525782012. View Article : Google Scholar
9	Turke AB, Zejnullahu K, Wu YL, Song Y, Dias-Santagata D, Lifshits E, Toschi L, Rogers A, Mok T, Sequist L, et al: Preexistence and clonal selection of MET amplification in EGFR mutant NSCLC. Cancer Cell. 17:77–88. 2010. View Article : Google Scholar
10	Debnath J and Brugge JS: Modelling glandular epithelial cancers in three-dimensional cultures. Nat Rev Cancer. 5:675–688. 2005. View Article : Google Scholar
11	Weigelt B and Bissell MJ: Unraveling the microenvironmental influences on the normal mammary gland and breast cancer. Semin Cancer Biol. 18:311–321. 2008. View Article : Google Scholar
12	Yamada KM and Cukierman E: Modeling tissue morphogenesis and cancer in 3D. Cell. 130:601–610. 2007. View Article : Google Scholar
13	Buick RN, Pullano R and Trent JM: Comparative properties of five human ovarian adenocarcinoma cell lines. Cancer Res. 45:3668–3676. 1985.
14	Appels NM, Beijnen JH and Schellens JH: Development of farnesyl transferase inhibitors: A review. Oncologist. 10:565–578. 2005. View Article : Google Scholar
15	Gschwendt M, Müller HJ, Kielbassa K, Zang R, Kittstein W, Rincke G and Marks F: Rottlerin, a novel protein kinase inhibitor. Biochem Biophys Res Commun. 199:93–98. 1994. View Article : Google Scholar
16	Scheipl S, Barnard M, Cottone L, Jorgensen M, Drewry DH, Zuercher WJ, Turlais F, Ye H, Leite AP, Smith JA, et al: EGFR inhibitors identified as a potential treatment for chordoma in a focused compound screen. J Pathol. 239:320–334. 2016. View Article : Google Scholar
17	Mishra R, Patel H, Alanazi S, Yuan L and Garrett JT: HER3 signaling and targeted therapy in cancer. Oncol Rev. 12:3552018.
18	Tabatabaei-Dakhili SA, Aguayo-Ortiz R, Domínguez L and Velázquez-Martínez CA: Untying the knot of transcription factor druggability: Molecular modeling study of FOXM1 inhibitors. J Mol Graph Model. 80:197–210. 2018. View Article : Google Scholar
19	Kalinichenko VV and Kalin TV: Is there potential to target FOXM1 for ‘undruggable’ lung cancers? Expert Opin Ther Targets. 19:865–867. 2015. View Article : Google Scholar
20	Trusolino L and Bertotti A: Compensatory pathways in oncogenic kinase signaling and resistance to targeted therapies: Six degrees of separation. Cancer Discov. 2:876–880. 2012. View Article : Google Scholar
21	Uprety D and Adjei AA: KRAS: From undruggable to a druggable cancer target. Cancer Treat Rev. 89:1020702020. View Article : Google Scholar
22	Elmariah H and DeZern AE: Chronic myelomonocytic leukemia: 2018 update to prognosis and treatment. Curr Hematol Malig Rep. 14:154–163. 2019. View Article : Google Scholar
23	Erba HP, Othus M, Walter RB, Kirschbaum MH, Tallman MS, Larson RA, Slovak ML, Kopecky KJ, Gundacker HM and Appelbaum FR: Four different regimens of farnesyltransferase inhibitor tipifarnib in older, untreated acute myeloid leukemia patients: North American Intergroup Phase II study SWOG S0432. Leuk Res. 38:329–333. 2014. View Article : Google Scholar
24	Yam C, Murthy RK, Valero V, Szklaruk J, Shroff GS, Stalzer CJ, Buzdar AU, Murray JL, Yang W, Hortobagyi GN, et al: A phase II study of tipifarnib and gemcitabine in metastatic breast cancer. Invest New Drugs. 36:299–306. 2018. View Article : Google Scholar
25	Vasan N, Boyer JL and Herbst RS: A RAS renaissance: Emerging targeted therapies for KRAS-mutated non-small cell lung cancer. Clin Cancer Res. 20:3921–30. 2014. View Article : Google Scholar
26	Giorgi C, Agnoletto C, Baldini C, Bononi A, Bonora M, Marchi S, Missiroli S, Patergnani S, Poletti F, Rimessi A, et al: Redox control of protein kinase C: Cell- and disease-specific aspects. Antioxid Redox Signal. 13:1051–1085. 2010. View Article : Google Scholar
27	Soltoff SP: Rottlerin: An inappropriate and ineffective inhibitor of PKCdelta. Trends Pharmacol Sci. 28:453–458. 2007. View Article : Google Scholar
28	Hynes NE and MacDonald G: ErbB receptors and signaling pathways in cancer. Curr Opin Cell Biol. 21:177–184. 2009. View Article : Google Scholar
29	Newby JC, Johnston SR, Smith IE and Dowsett M: Expression of epidermal growth factor receptor and c-erbB2 during the development of tamoxifen resistance in human breast cancer. Clin Cancer Res. 3:1643–1651. 1997.
30	Shou J, Massarweh S, Osborne CK, Wakeling AE, Ali S, Weiss H and Schiff R: Mechanisms of tamoxifen resistance: Increased estrogen receptor-HER2/neu cross-talk in ER/HER2-positive breast cancer. J Natl Cancer Inst. 96:926–935. 2004. View Article : Google Scholar