Caglar BERKEL, Burak KUCUK, Merve USTA, Esra YILMAZ, Ercan CACAN

The Effect of Olaparib and Bortezomib Combination Treatment on Ovarian Cancer Cell Lines

Objective: Ovarian cancer (OC) is the deadliest gynecologic malignancy and has a poor survival rate due to late diagnosis and chemoresistance development. In the standard treatment of OC, platinum-based chemotherapeutics are used. However, following several rounds of chemotherapy, these drugs’ efficacy eventually becomes limited due to the chemoresistance in most patients who previously responded to this treatment. Therefore, overcoming chemoresistance in the treatment of OC is of high importance. In this study, we investigated the effect of combinatorial inhibition of poly(ADP-ribose)polymerase (PARP) and proteasome by olaparib and bortezomib on chemosensitive and chemoresistant OC cell lines. Materials and Methods: We used sulphorhodamine B assay to screen cell viability following drug treatments alone or in combination, and used the cytotoxicity data to model the effect of drugs on cell death in R programming environment. In addition to olaparib and bortezomib, we performed cytotoxicity screenings where we applied cisplatin to OC cells. We also carried out flow cytometry analysis to quantify apoptotic cells following treatments. Results: We showed that combination treatment was more effective on chemosensitive OC cell lines when cisplatin was not used. In the presence of cisplatin, olaparib and bortezomib combination treatment resulted in higher cytotoxicity in chemoresistant OC lines compared to chemosensitive OC cell lines. Combinatorial inhibition of PARP and proteasome led to a higher number of apoptotic cells in OV2008 chemosensitive cell line compared to drugs alone. Conclusion: Our data shows that olaparib and bortezomib combination treatment might show promise in vivo in the treatment of OC. Also, the efficacy of this combination treatment might be dependent on OC cells’ chemosensitivity profiles.

Keywords:

Ovarian cancer, olaparib, bortezomib, chemoresistance, proteasome,

PDF

___

1 Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2018;68(6):394-424.
2 Berkel C, Cacan E. In silico analysis of DYNLL1 expression in ovarian cancer chemoresistance. Cell Biol Int 2020;44(8):1598-605.
3 Ali MW, Cacan E, Liu Y, Pierce JY, Creasman WT, Murph MM, Govindarajan R, Eblen ST, Greer SF, Hooks SB. Transcriptional suppression, DNA methylation, and histone deacetylation of the regulator of G-protein signaling 10 (RGS10) gene in ovarian cancer cells. PLoS One 2013;8(3):e60185.
4 Cacan E, Ali MW, Boyd NH, Hooks SB, Greer SF. Inhibition of HDAC1 and DNMT1 modulate RGS10 expression and decrease ovarian cancer chemoresistance. PLoS One 2014;9(1): e87455.
5 European Medicines Agency. Lynparza summary of product characteristics, 2018; Available at: http://ec.europa.eu/health/documents/community-register/2018/20180508140545/anx_140545_en.pdf.
6 Morales J, Li L, Fattah FJ, Dong Y, Bey EA, Patel M, et al. Review of poly (ADP-ribose) polymerase (PARP) mechanisms of action and rationale for targeting in cancer and other diseases. Crit Rev Eukaryot Gene Expr 2014; 24(1):15-28.
7 Vescarelli E, Gerini G, Megiorni F, Anastasiadou E, Pontecorvi P, Solito L, et al. MiR-200c sensitizes Olaparib-resistant ovarian cancer cells by targeting Neuropilin 1. J Exp Clin Cancer Res 2020; 39(1):3.
8 Wang L, Shi C, Wright FA, Guo D, Wang X, Wang D, et al. Multifunctional telodendrimer nanocarriers restore synergy of bortezomib and doxorubicin in ovarian cancer treatment. Cancer Res 2017; 77(12):3293-305.
9 Cacan E, Spring AM, Kumari A, Greer SF, Garnett-Benson C. Combination treatment with sublethal ionizing radiation and the proteasome inhibitor, bortezomib, enhances death-receptor mediated apoptosis and anti-tumor immune attack. Int J Mol Sci 2015; 16(12):30405-21.
10 Takenaka M, Saito M, Iwakawa R, Yanaihara N, Saito M, Kato M, et al. Profiling of actionable gene alterations in ovarian cancer by targeted deep sequencing. Int J Oncol 2015; 46(6):2389-98.
11 Anglesio MS, Wiegand KC, Melnyk N, Chow C, Salamanca C, Prentice LM, et al. Type-specific cell line models for type-specific ovarian cancer research. PLoS One 2013; 8(9):e72162.
12 Beaufort CM, Helmijr JC, Piskorz AM, Hoogstraat M, Ruigrok-Ritstier K, Besselink N, et al. Ovarian cancer cell line panel (OCCP): clinical importance of in vitro morphological subtypes. PLoS One 2014; 9(9):e103988.
13 Vichai V, Kirtikara K. Sulforhodamine B colorimetric assay for cytotoxicity screening. Nat Protoc 2006; 1(3):1112-6.
14 Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, et al. New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 1990; 82(13):1107-112.
15 R Core Team. R: A language and environment for statistical computing. R Foundation for Statistical Computing, 2018; Vienna, Austria. URL https://www.R-project.org/.
16 Grolemund G, Wickham H. R for data science, 2016; 1st edition. California: O’Reilly.
17 Holmes S, Huber W. Modern statistics for modern biology, 2019; 1st edition. Cambridge: Cambridge University Press.
18 Wickham H. tidyverse: Easily Install and Load the ‘Tidyverse’. R package version, 2017, 1.2.1. https://CRAN.R-project.org/package=tidyverse
19 Wickham H and Bryan J. readxl: Read Excel Files. R package version 1.3.1, 2019. https://CRAN.R-project.org/package=readxl
20 Soetaert K. plot3D: Plotting Multi-Dimensional Data. R package version 1.3, 2019. https://CRAN.R-project.org/package=plot3D
21 Ooms J. magick: Advanced graphics and image-processing in R. R package version 2.4.0, 2020. https://CRAN.R-project.org/package=magick
22 Auguie B. gridExtra: Miscellaneous functions for "Grid" graphics. R package version 2.3, 2017. https://CRAN.R-project.org/package=gridExtra
23 Kassambara A. ggpubr: 'ggplot2' Based Publication Ready Plots. R package version 0.4.0, 2020. https://CRAN.R-project.org/package=ggpubr
24 Xie Y, Allaire JJ, Grolemund G. R Markdown: The Definitive Guide. Chapman and Hall/CRC, 2018. ISBN 9781138359338. URL https://bookdown.org/yihui/rmarkdown.
25 Xie Y, Dervieux C, Riederer E. R Markdown Cookbook. Chapman and Hall/CRC, 2020. ISBN 9780367563837. URL https://bookdown.org/yihui/rmarkdown-cookbook.
26 He YJ, Meghani K, Caron MC, Yang C, Ronato DA, Bian J, et al. DYNLL1 binds to MRE11 to limit DNA end resection in BRCA1-deficient cells. Nature 2018;563(7732):522-6.
27 Janyst K, Janyst M, Siernicka M, Lasek W. Synergistic antitumor effects of histone deacetylase inhibitor scriptaid and bortezomib against ovarian cancer cells. Oncol Rep 2018;39(4):1999-2005.
28 Wang L, Shi C, Wright FA, Guo D, Wang X, Wang D, et al. Multifunctional telodendrimer nanocarriers restore synergy of bortezomib and doxorubicin in ovarian cancer treatment. Cancer Res 2017;77(12):3293-305.
29 Berkel C, Cacan E. GAB2 and GAB3 are expressed in a tumor stage-, grade- and histotype-dependent manner and are associated with shorter progression-free survival in ovarian cancer. J Cell Commun Signal 2020;10.1007/s12079-020-00582-3.
30 Berkel C, Cacan E. Single-cell epigenomics in cancer research. Biomed J Sci&Tech Res 2019;21(3).
31 Kroeger PT, Jr, & Drapkin R. Pathogenesis and heterogeneity of ovarian cancer. Curr Opin Obstet Gynecol 2017; 29(1), 26–34.
32 Winterhoff BJ, Maile M, Mitra AK, Sebe A, Bazzaro M, Geller MA, et al. Single cell sequencing reveals heterogeneity within ovarian cancer epithelium and cancer associated stromal cells. Gynecol Oncol 2017;144(3):598-606.
33 Shih AJ, Menzin A, Whyte J, Lovecchio J, Liew A, Khalili H, et al. Identification of grade and origin specific cell populations in serous epithelial ovarian cancer by single cell RNA-seq. PLoS One 2018;13(11):e0206785.