Destabilization of FoxM1 and Inhibition of Topoisomerase I Contribute to Cytotoxicity of Prenylated Xanthones Isolated from Metaxya rostrata ^{[ # ]}

Abstract

We recently isolated the prenylated xanthones 2-deprenyl-rheediaxanthone B (XB) and 2-deprenyl-7-hydroxy-rheediaxanthone B (OH-XB) from the South American tree fern Metaxya rostrata. This study explores the mechanisms underlying the FoxM1 downregulation induced by both xanthones. Analysis of cell viability and cell-death induction in SW480, HCT116, Caco-2, DLD1 and HT29 exposed to xanthones found cell-loss and activation of caspase in all cell lines except HT29 that do not have high FoxM1 protein levels. To determine the cellular mechanism of xanthone-induced FoxM1 loss, protein stability was analyzed by cycloheximide-chase experiments and showed reduction of FoxM1 stability by XB but not OH-XB. Destabilization was prevented by inhibiting proteasome activity using MG-132 and moderately by the lysosomal inhibitor bafilomycin A1 (baf A1). OH-XB had a stronger impact than XB on FoxM1 mRNA expression by qRT-PCR, and MG-132 positively affected FoxM1 protein level in OH-XB exposed cells even though no decrease in protein abundance had been induced by the xanthone. Additionally, the compound inhibited topoisomerase I causing DNA DSB and early cell cycle arrest. This may reduce FoxM1 gene expression, which may in turn compromise DNA repair and enhance xanthone-induced cell death. With regard to xanthone-induced cell death, MG-132 protected cultures from cell loss induced by both compounds, and baf A1 was active against these XB-induced effects. In summary, both destabilization of FoxM1 protein and topoisomerase I inhibition contribute to both XB and OH-XB cytotoxic activity albeit at different ratios.

Key words

2-deprenyl-rheediaxanthone B, 2-deprenyl-7-hydroxy-rheediaxanthone B, Metaxya rostrata - Metaxyaceae - FoxM1 - topoisomerase I

• References

• 1 Newman DJ, Cragg GM. Natural products as sources of new drugs from 1981 to 2014. J Nat Prod 2016; 79: 629-661
• 2 Atanasov AG, Waltenberger B, Pferschy-Wenzig EM, Linder T, Wawrosch C, Uhrin P, Temml V, Wang L, Schwaiger S, Heiss EH, Rollinger JM, Schuster D, Breuss JM, Bochkov V, Mihovilovic MD, Kopp B, Bauer R, Dirsch VM, Stuppner H. Discovery and resupply of pharmacologically active plant-derived natural products: a review. Biotechnol Adv 2015; 33: 1582-1614
• 3 Castanheiro RA, Silva AM, Campos NA, Nascimento MS, Pinto MM. Antitumor activity of some prenylated xanthones. Pharmaceuticals (Basel) 2009; 2: 33-43
• 4 Ding L, Liu B, Qi LL, Zhou QY, Hou Q, Li J, Zhang Q. Anti-proliferation, cell cycle arrest and apoptosis induced by a natural xanthone from Gentianopsis paludosa Ma, in human promyelocytic leukemia cell line HL-60 cells. Toxicol In Vitro 2009; 23: 408-417
• 5 Matsumoto K, Akao Y, Yi H, Ohguchi K, Ito T, Tanaka T, Kobayashi E, Iinuma M, Nozawa Y. Preferential target is mitochondria in alpha-mangostin-induced apoptosis in human leukemia HL60 cells. Bioorg Med Chem 2004; 12: 5799-5806
• 6 Watanapokasin R, Jarinthanan F, Nakamura Y, Sawasjirakij N, Jaratrungtawee A, Suksamrarn S. Effects of alpha-mangostin on apoptosis induction of human colon cancer. World J Gastroenterol 2011; 17: 2086-2095
• 7 Mittermair E, Krenn L, Marian B. Prenylated xanthones from Metaxya rostrata suppress FoxM1 and induce active cell death by distinct mechanisms. Phytomedicine 2019; 60: 152912
• 8 Kainz KP, Krenn L, Erdem Z, Kaehlig H, Zehl M, Bursch W, Berger W, Marian B. 2-deprenyl-rheediaxanthone B isolated from Metaxya rostrata induces active cell death in colorectal tumor cells. PLoS One 2013; 8: e65745
• 9 Wierstra I. FOXM1 (Forkhead box M1) in tumorigenesis: overexpression in human cancer, implication in tumorigenesis, oncogenic functions, tumor-suppressive properties, and target of anticancer therapy. Adv Cancer Res 2013; 119: 191-419
• 10 Costa RH. FoxM1 dances with mitosis. Nat Cell Biol 2005; 7: 108-110
• 11 Liu M, Dai B, Kang SH, Ban K, Huang FJ, Lang FF, Aldape KD, Xie TX, Pelloski CE, Xie K, Sawaya R, Huang S. FoxM1B is overexpressed in human glioblastomas and critically regulates the tumorigenicity of glioma cells. Cancer Res 2006; 66: 3593-3602
• 12 Liu Y, Hock JM, Van Beneden RJ, Li X. Aberrant overexpression of FOXM1 transcription factor plays a critical role in lung carcinogenesis induced by low doses of arsenic. Mol Carcinog 2014; 53: 380-391
• 13 Anders L, Ke N, Hydbring P, Choi YJ, Widlund HR, Chick JM, Zhai H, Vidal M, Gygi SP, Braun P, Sicinski P. A systematic screen for CDK4/6 substrates links FOXM1 phosphorylation to senescence suppression in cancer cells. Cancer Cell 2011; 20: 620-634
• 14 Li SK, Smith DK, Leung WY, Cheung AM, Lam EW, Dimri GP, Yao KM. FoxM1c counteracts oxidative stress-induced senescence and stimulates Bmi-1 expression. J Biol Chem 2008; 283: 16545-16553
• 15 Wierstra I. The transcription factor FOXM1 (Forkhead box M1): proliferation-specific expression, transcription factor function, target genes, mouse models, and normal biological roles. Adv Cancer Res 2013; 118: 97-398
• 16 Alvarez-Fernandez M, Halim VA, Aprelia M, Laoukili J, Mohammed S, Medema RH. Protein phosphatase 2A (B55alpha) prevents premature activation of forkhead transcription factor FoxM1 by antagonizing cyclin A/cyclin-dependent kinase-mediated phosphorylation. J Biol Chem 2011; 286: 33029-33036
• 17 Halasi M, Gartel AL. Targeting FOXM1 in cancer. Biochem Pharmacol 2013; 85: 644-652
• 18 Xie Z, Tan G, Ding M, Dong D, Chen T, Meng X, Huang X, Tan Y. Foxm1 transcription factor is required for maintenance of pluripotency of P19 embryonal carcinoma cells. Nucleic Acids Res 2010; 38: 8027-8038
• 19 Laoukili J, Alvarez M, Meijer LA, Stahl M, Mohammed S, Kleij L, Heck AJ, Medema RH. Activation of FoxM1 during G2 requires cyclin A/Cdk-dependent relief of autorepression by the FoxM1 N-terminal domain. Mol Cell Biol 2008; 28: 3076-3087
• 20 Laoukili J, Kooistra MR, Bras A, Kauw J, Kerkhoven RM, Morrison A, Clevers H, Medema RH. FoxM1 is required for execution of the mitotic programme and chromosome stability. Nat Cell Biol 2005; 7: 126-136
• 21 Laoukili J, Stahl M, Medema RH. FoxM1: at the crossroads of ageing and cancer. Biochim Biophys Acta 2007; 1775: 92-102
• 22 Park HJ, Wang Z, Costa RH, Tyner A, Lau LF, Raychaudhuri P. An N-terminal inhibitory domain modulates activity of FoxM1 during cell cycle. Oncogene 2008; 27: 1696-1704
• 23 Buchanan BW, Lloyd ME, Engle SM, Rubenstein EM. Cycloheximide chase analysis of protein degradation in Saccharomyces cerevisiae . J Vis Exp 2016; 110: e53975
• 24 Staker BL, Feese MD, Cushman M, Pommier Y, Zembower D, Stewart L, Burgin AB. Structures of 3 classes of anticancer agents bound to the human topoisomerase I-DNA covalent complex. J Med Chem 2005; 48: 2336-2345
• 25 Champoux JJ, Dulbecco R. An activity from mammalian cells that untwists superhelical DNA – a possible swivel for DNA replication (polyoma-ethidium bromide-mouse-embryo cells-dye binding assay). Proc Natl Acad Sci U S A 1972; 69: 143-146
• 26 Gonzalez-Ruiz V, Pascua I, Fernandez-Marcelo T, Ribelles P, Bianchini G, Sridharan V, Iniesta P, Ramos MT, Olives AI, Martin MA, Menendez JC. B-ring-aryl substituted luotonin A analogues with a new binding mode to the topoisomerase 1-DNA complex show enhanced cytotoxic activity. PLoS One 2014; 9: e95998
• 27 Nitiss JL, Soans E, Rogojina A, Seth A, Mishina M. Topoisomerase assays. Curr Protoc Pharmacol 2012; Chapter 3: Unit3.3
• 28 Hsiang YH, Lihou MG, Liu LF. Arrest of replication forks by drug-stabilized topoisomerase I-DNA cleavable complexes as a mechanism of cell killing by camptothecin. Cancer Res 1989; 49: 5077-5082
• 29 Tsao YP, Russo A, Nyamuswa G, Silber R, Liu LF. Interaction between replication forks and topoisomerase I-DNA cleavable complexes: studies in a cell-free SV40 DNA replication system. Cancer Res 1993; 53: 5908-5914
• 30 Monteiro LJ, Khongkow P, Kongsema M, Morris JR, Man C, Weekes D, Koo CY, Gomes AR, Pinto PH, Varghese V, Kenny LM, Charles Coombes R, Freire R, Medema RH, Lam EW. The Forkhead Box M1 protein regulates BRIP1 expression and DNA damage repair in epirubicin treatment. Oncogene 2013; 32: 4634-4645
• 31 Park YY, Jung SY, Jennings NB, Rodriguez-Aguayo C, Peng G, Lee SR, Kim SB, Kim K, Leem SH, Lin SY, Lopez-Berestein G, Sood AK, Lee JS. FOXM1 mediates Dox resistance in breast cancer by enhancing DNA repair. Carcinogenesis 2012; 33: 1843-1853
• 32 Koo CY, Muir KW, Lam EW. FOXM1: From cancer initiation to progression and treatment. Biochim Biophys Acta 2012; 1819: 28-37
• 33 Raychaudhuri P, Park HJ. FoxM1: a master regulator of tumor metastasis. Cancer Res 2011; 71: 4329-4333
• 34 Teh MT. FOXM1 coming of age: time for translation into clinical benefits?. Front Oncol 2012; 2: 146
• 35 Wilson MS, Brosens JJ, Schwenen HD, Lam EW. FOXO and FOXM1 in cancer: the FOXO-FOXM1 axis shapes the outcome of cancer chemotherapy. Curr Drug Targets 2011; 12: 1256-1266
• 36 Zhao F, Lam EW. Role of the forkhead transcription factor FOXO-FOXM1 axis in cancer and drug resistance. Front Med 2012; 6: 376-380
• 37 Yu L, Chen Y, Tooze SA. Autophagy pathway: Cellular and molecular mechanisms. Autophagy 2018; 14: 207-215
• 38 Wu WK, Sung JJ, Wu YC, Li ZJ, Yu L, Cho CH. Bone morphogenetic protein signalling is required for the anti-mitogenic effect of the proteasome inhibitor MG-132 on colon cancer cells. Br J Pharmacol 2008; 154: 632-638
• 39 Delgado JL, Hsieh CM, Chan NL, Hiasa H. Topoisomerases as anticancer targets. Biochem J 2018; 475: 373-398
• 40 Erdem ZN, Schwarz S, Drev D, Heinzle C, Reti A, Heffeter P, Hudec X, Holzmann K, Grasl-Kraupp B, Berger W, Grusch M, Marian B. Irinotecan upregulates fibroblast growth factor receptor 3 expression in colorectal cancer cells, which mitigates irinotecan-induced apoptosis. Transl Oncol 2017; 10: 332-339
• 41 Sonvilla G, Allerstorfer S, Heinzle C, Stattner S, Karner J, Klimpfinger M, Wrba F, Fischer H, Gauglhofer C, Spiegl-Kreinecker S, Grasl-Kraupp B, Holzmann K, Grusch M, Berger W, Marian B. Fibroblast growth factor receptor 3-IIIc mediates colorectal cancer growth and migration. Br J Cancer 2010; 102: 1145-1156