Abstract
Monastrol is an allosteric inhibitor of the mitotic kinesin Eg5 that exhibits an antiproliferative effect against several cell lines. We investigated the antiproliferative effect of monastrol on human breast adenocarcinoma cells (MCF-7) and mammary epithelial cells (HB4a, non-tumoral). Monastrol treatment decreased cell viability only in MCF-7 tumor cells. Real-time cell growth kinetic analysis showed a decrease in the proliferation of MCF-7 cells exposed to monastrol, while in the HB4a cells, only a concentration of 100 μM was able to induce this effect. In a cell cycle analysis, exposure of MCF-7 cells to monastrol led to an increased population of cells in both the G1 and G2/M phases. In HB4a cells, the proportion of cells in the G2/M phase was increased. Monastrol led to an increased mitotic index in both cell lines. Monastrol was not able to induce cell death by apoptosis in any of the cell lines studied. Gene expression analysis was performed to measure the mRNA levels of cell cycle genes, DNA damage indicator gene, and apoptotic related genes. Treatment with monastrol induced in MCF-7 cells a 5-fold increase in the mRNA levels of the CDKN1A gene, an inhibitor of CDKs related with cell cycle arrest in response a stress stimulus, and a 2-fold decrease in CDKN1C mRNA levels in HB4a cells. These results provide evidence that monastrol has a greater antiproliferative effect on MCF-7 tumor cells compared with non-tumor HB4a cells; however, no selective is observed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abassi YA, ** B, Zhang W, et al. (2009) Kinetic cell-based morphological screening: prediction of mechanism of compound action and off-target effects. Chem Biol 16:712–723. doi:10.1016/j.chembiol.2009.05.011
Abbas T, Dutta A (2009) p21 in cancer: intricate networks and multiple activities. Nat Rev Cancer 9:400–414. doi:10.1038/nrc2657
Abukhdeir AM, Park BH (2008) P21 and p27: roles in carcinogenesis and drug resistance. Expert Rev Mol Med 10:e19. doi:10.1017/S1462399408000744
Asraf H, Amsterdam A, Ben-Menahem D (2015) Modulation of the steroidogenic related activity according to the design of single-chain bovine FSH analogs. Gen Comp Endocrinol 216:171–181. doi:10.1016/j.ygcen.2015.04.004
Blangy A, Lane HA, d’Herin P, et al. (1995) Phosphorylation by p34cdc2 regulates spindle association of human Eg5, a kinesin-related motor essential for bipolar spindle formation in vivo. Cell 83:1159–1169
Chin GM, Herbst R (2006) Induction of apoptosis by monastrol, an inhibitor of the mitotic kinesin Eg5, is independent of the spindle checkpoint. Mol Cancer Ther 5:2580–2591. doi:10.1158/1535-7163.MCT-06-0201
Costa-Rosa Md (2011) Interferindo na progressão do ciclo celular para avaliar possíveis alterções de ploidia em células tumorais de mam humana. Universidade de São Paulo
DeBonis S, Simorre JP, Crevel I, et al. (2003) Interaction of the mitotic inhibitor monastrol with human kinesin Eg5. Biochemistry 42:338–349. doi:10.1021/bi026716j
Enos AP, Morris NR (1990) Mutation of a gene that encodes a kinesin-like protein blocks nuclear division in A. nidulans. Cell 60:1019–1027
Ferenz NP, Gable A, Wadsworth P (2010) Mitotic functions of kinesin-5. Semin Cell Dev Biol 21:255–259. doi:10.1016/j.semcdb.2010.01.019
Ferlay J, Soerjomataram I, Dikshit R, et al. (2015) Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 136:E359–E386. doi:10.1002/ijc.29210
Gascoigne KE, Taylor SS (2008) Cancer cells display profound intra- and interline variation following prolonged exposure to antimitotic drugs. Cancer Cell 14:111–122. doi:10.1016/j.ccr.2008.07.002
Green DR, Kroemer G (2009) Cytoplasmic functions of the tumour suppressor p53. Nature 458:1127–1130. doi:10.1038/nature07986
Hanahan D, Weinberg RA (2000) The hallmarks of cancer. Cell 100:57–70
Hanahan D, Weinberg RA (2011) Hallmarks of cancer: the next generation. Cell 144:646–674. doi:10.1016/j.cell.2011.02.013
Haque SA, Hasaka TP, Brooks AD, et al. (2004) Monastrol, a prototype anti-cancer drug that inhibits a mitotic kinesin, induces rapid bursts of axonal outgrowth from cultured postmitotic neurons. Cell Motil Cytoskeleton 58:10–16. doi:10.1002/cm.10176
Harrison MR, Holen KD, Liu G (2009) Beyond taxanes: a review of novel agents that target mitotic tubulin and microtubules, kinases, and kinesins. Clin Adv Hematol Oncol 7:54–64
Huszar D, Theoclitou ME, Skolnik J, Herbst R (2009) Kinesin motor proteins as targets for cancer therapy. Cancer Metastasis Rev 28:197–208. doi:10.1007/s10555-009-9185-8
Kapoor TM, Mayer TU, Coughlin ML, Mitchison TJ (2000) Probing spindle assembly mechanisms with monastrol, a small molecule inhibitor of the mitotic kinesin, Eg5. J Cell Biol 150:975–988
Kaur J, Sundar S, Singh N (2010) Molecular docking, structure-activity relationship and biological evaluation of the anticancer drug monastrol as a pteridine reductase inhibitor in a clinical isolate of Leishmania donovani. J Antimicrob Chemother 65:1742–1748. doi:10.1093/jac/dkq189
Kavanagh E, Joseph B (2011) The hallmarks of CDKN1C (p57, KIP2) in cancer. Biochim Biophys Acta 1816:50–56. doi:10.1016/j.bbcan.2011.03.002
Koller E, Propp S, Zhang H, et al. (2006) Use of a chemically modified antisense oligonucleotide library to identify and validate Eg5 (kinesin-like 1) as a target for antineoplastic drug development. Cancer Res 66:2059–2066. doi:10.1158/0008-5472.CAN-05-1531
Larson PS, Schlechter BL, King CL, et al. (2008) CDKN1C/p57kip2 is a candidate tumor suppressor gene in human breast cancer. BMC Cancer 8:68. doi:10.1186/1471-2407-8-68
Le Guellec R, Paris J, Couturier A, et al. (1991) Cloning by differential screening of a Xenopus cDNA that encodes a kinesin-related protein. Mol Cell Biol 11:3395–3398
Leizerman I, Avunie-Masala R, Elkabets M, et al. (2004) Differential effects of monastrol in two human cell lines. Cell Mol Life Sci 61:2060–2070. doi:10.1007/s00018-004-4074-3
Lin S, Liu M, Son YJ, et al. (2011) Inhibition of Kinesin-5, a microtubule-based motor protein, as a strategy for enhancing regeneration of adult axons. Traffic 12:269–286. doi:10.1111/j.1600-0854.2010.01152.x
Maliga Z, Mitchison TJ (2006) Small-molecule and mutational analysis of allosteric Eg5 inhibition by monastrol. BMC Chem Biol 6:2. doi:10.1186/1472-6769-6-2
Maliga Z, Kapoor TM, Mitchison TJ (2002) Evidence that monastrol is an allosteric inhibitor of the mitotic kinesin Eg5. Chem Biol 9:989–996
Mandal M, Bandyopadhyay D, Goepfert TM, Kumar R (1998) Interferon-induces expression of cyclin-dependent kinase-inhibitors p21WAF1 and p27Kip1 that prevent activation of cyclin-dependent kinase by CDK-activating kinase (CAK. Oncogene 16:217–225. doi:10.1038/sj.onc.1201529
Mayer TU, Kapoor TM, Haggarty SJ, et al. (1999) Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen. Science 286:971–974
Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63
Pfaffl MW, Horgan GW, Dempfle L (2002) Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 30:e36
Rieder CL, Maiato H (2004) Stuck in division or passing through: what happens when cells cannot satisfy the spindle assembly checkpoint. Dev Cell 7:637–651. doi:10.1016/j.devcel.2004.09.002
Rowinsky EK, Chaudhry V, Cornblath DR, Donehower RC (1993) Neurotoxicity of Taxol. J Natl Cancer Inst Monogr:107–115
Russowsky D, Canto RF, Sanches SA, et al. (2006) Synthesis and differential antiproliferative activity of Biginelli compounds against cancer cell lines: monastrol, oxo-monastrol and oxygenated analogues. Bioorg Chem 34:173–182. doi:10.1016/j.bioorg.2006.04.003
Sakowicz R, Finer JT, Beraud C, et al. (2004) Antitumor activity of a kinesin inhibitor. Cancer Res 64:3276–3280
Savio AL, da Silva GN, de Camargo EA, Salvadori DM (2014) Cell cycle kinetics, apoptosis rates, DNA damage and TP53 gene expression in bladder cancer cells treated with allyl isothiocyanate (mustard essential oil. Mutat Res Fundam Mol Mech Mutagen 762:40–46. doi:10.1016/j.mrfmmm.2014.02.006
Sherr CJ, Roberts JM (1999) CDK inhibitors: positive and negative regulators of G1-phase progression. Genes Dev 13:1501–1512
Shibata MA, Yoshidome K, Shibata E, et al. (2001) Suppression of mammary carcinoma growth in vitro and in vivo by inducible expression of the Cdk inhibitor p21. Cancer Gene Ther 8:23–35. doi:10.1038/sj.cgt.7700275
Thompson SL, Compton DA (2008) Examining the link between chromosomal instability and aneuploidy in human cells. J Cell Biol 180:665–672. doi:10.1083/jcb.200712029
Thompson SL, Compton DA (2010) Proliferation of aneuploid human cells is limited by a p53-dependent mechanism. J Cell Biol 188:369–381. doi:10.1083/jcb.200905057
Tuxen MK, Hansen SW (1994) Neurotoxicity secondary to antineoplastic drugs. Cancer Treat Rev 20:191–214
Ullah Z, Kohn MJ, Yagi R, et al. (2008) Differentiation of trophoblast stem cells into giant cells is triggered by p57/Kip2 inhibition of CDK1 activity. Genes Dev 22:3024–3036. doi:10.1101/gad.1718108
Vijapurkar U, Wang W, Herbst R (2007) Potentiation of kinesin spindle protein inhibitor-induced cell death by modulation of mitochondrial and death receptor apoptotic pathways. Cancer Res 67:237–245. doi:10.1158/0008-5472.CAN-06-2406
Weaver BA, Cleveland DW (2005) Decoding the links between mitosis, cancer, and chemotherapy: the mitotic checkpoint, adaptation, and cell death. Cancer Cell 8:7–12. doi:10.1016/j.ccr.2005.06.011
Xu XY, Wang WQ, Zhang L, et al. (2012) Clinical implications of p57 KIP2 expression in breast cancer. Asian Pac J Cancer Prev 13:5033–5036
Xu C, Klaw MC, Lemay MA, et al. (2015) Pharmacologically inhibiting kinesin-5 activity with monastrol promotes axonal regeneration following spinal cord injury. Exp Neurol 263:172–176. doi:10.1016/j.expneurol.2014.10.013
Yoon SY, Choi JE, Huh JW, et al. (2005) Monastrol, a selective inhibitor of the mitotic kinesin Eg5, induces a distinctive growth profile of dendrites and axons in primary cortical neuron cultures. Cell Motil Cytoskeleton 60:181–190. doi:10.1002/cm.20057
Acknowledgments
This research was support by CNPq, CAPES, and Fundação Araucária, Brazil.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
ESM 1
(DOCX 34 kb)
Rights and permissions
About this article
Cite this article
Marques, L.A., Semprebon, S.C., Niwa, A.M. et al. Antiproliferative activity of monastrol in human adenocarcinoma (MCF-7) and non-tumor (HB4a) breast cells. Naunyn-Schmiedeberg's Arch Pharmacol 389, 1279–1288 (2016). https://doi.org/10.1007/s00210-016-1292-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00210-016-1292-9