title: Targeting mitotic exit with hyperthermia or APC/C inhibition to increase paclitaxel efficacy.
abstract: Microtubule-poisoning drugs, such as Paclitaxel (or Taxol, PTX), are powerful and commonly used anti-neoplastic agents for the treatment of several malignancies. PTX triggers cell death, mainly through a mitotic arrest following the activation of the spindle assembly checkpoint (SAC). Cells treated with PTX slowly slip from this mitotic block and die by mitotic catastrophe. However, cancer cells can acquire or are intrinsically resistant to this drug, posing one of the main obstacles for PTX clinical effectiveness. In order to override PTX resistance and increase its efficacy, we investigated both the enhancement of mitotic slippage and the block of mitotic exit. To test these opposing strategies, we used physiological hyperthermia (HT) to force exit from PTX-induced mitotic block and the anaphase-promoting complex/cyclosome (APC/C) inhibitor, proTAME, to block mitotic exit. We observed that application of HT on PTX-treated cells forced mitotic slippage, as shown by the rapid decline of cyclin B levels and by microscopy analysis. Similarly, HT induced mitotic exit in cells blocked in mitosis by other antimitotic drugs, such as Nocodazole and the Aurora A inhibitor MLN8054, indicating a common effect of HT on mitotic cells. On the other hand, proTAME prevented mitotic exit of PTX and MLN8054 arrested cells, prolonged mitosis, and induced apoptosis. In addition, we showed that proTAME prevented HT-mediated mitotic exit, indicating that stress-induced APC/C activation is necessary for HT-induced mitotic slippage. Finally, HT significantly increased PTX cytotoxicity, regardless of cancer cells' sensitivity to PTX, and this activity was superior to the combination of PTX with pro-TAME. Our data suggested that forced mitotic exit of cells arrested in mitosis by anti-mitotic drugs, such as PTX, can be a more successful anticancer strategy than blocking mitotic exit by inactivation of the APC/C.
authors.affiliation: Department of Anatomy and Cell Biology, University of Florida College of Medicine, Gainesville, FL, USA.
authors.forename: Serena
authors.initials: S
authors.lastname: Giovinazzi
authors.forename: Dhruv
authors.initials: D
authors.lastname: Bellapu
authors.forename: Viacheslav M
authors.initials: VM
authors.lastname: Morozov
authors.forename: Alexander M
authors.initials: AM
authors.lastname: Ishov
mesh_terms: D064173:Anaphase-Promoting Complex-Cyclosome / Q000037:antagonists & inhibitors*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D015153:Blotting, Western; D019926:Cyclin B / Q000378:metabolism; D006801:Humans; D006979:Hyperthermia, Induced / Q000379:methods*; D008938:Mitosis / Q000187:drug effects / Q000502:physiology*; D009369:Neoplasms / Q000188:drug therapy*; D017239:Paclitaxel / Q000494:pharmacology*
chemical_list: D000970:Antineoplastic Agents; D019926:Cyclin B; D064173:Anaphase-Promoting Complex-Cyclosome; D017239:Paclitaxel
keywords: APC/C; antimitotic drug resistance; aurora A inhibitors; hyperthermia; mitotic block; mitotic catastrophe; proTAME; taxanes
references.citation: Lancet Oncol. 2002 Aug;3(8):487-97
references.pmid: 12147435
references.citation: Oncologist. 2004;9 Suppl 2:24-32
references.pmid: 15161988
references.citation: Nature. 2011 Mar 3;471(7336):110-4
references.pmid: 21368834
references.citation: Eur J Cancer. 2010 Aug;46(12):2206-15
references.pmid: 20471824
references.citation: Cell Death Differ. 2012 Jan;19(1):107-20
references.pmid: 21760595
references.citation: Cancer Res. 2011 Jul 1;71(13):4608-16
references.pmid: 21712408
references.citation: Clin Cancer Res. 2010 Jan 15;16(2):566-76
references.pmid: 20068098
references.citation: Ann Oncol. 2002 Aug;13(8):1173-84
references.pmid: 12181239
references.citation: Nat Rev Cancer. 2004 Apr;4(4):253-65
references.pmid: 15057285
references.citation: Cancer Cell. 2009 Oct 6;16(4):274-5
references.pmid: 19800571
references.citation: Cancer Cell. 2010 Oct 19;18(4):382-95
references.pmid: 20951947
references.citation: Cell Cycle. 2009 May 15;8(10):1544-51
references.pmid: 19372739
references.citation: Eur J Cancer. 1999 Oct;35(10):1431-9
references.pmid: 10673974
references.citation: Cancer Chemother Pharmacol. 2005 Dec;56(6):615-22
references.pmid: 16025289
references.citation: Cell Div. 2012 Feb 09;7:4
references.pmid: 22321970
references.citation: Clin Cancer Res. 2000 Dec;6(12):4610-7
references.pmid: 11156210
references.citation: Curr Biol. 2006 Jun 20;16(12):1194-200
references.pmid: 16782009
references.citation: Drug Resist Updat. 2001 Oct;4(5):303-13
references.pmid: 11991684
references.citation: Oncogene. 2011 Jun 23;30(25):2799-809
references.pmid: 21339734
references.citation: Ann Surg Oncol. 2007 Mar;14(3):1220-8
references.pmid: 17206477
references.citation: J Cell Sci. 2009 Aug 1;122(Pt 15):2579-85
references.pmid: 19625502
references.citation: Cell Cycle. 2012 Jan 15;11(2):310-21
references.pmid: 22214763
references.citation: Mol Cancer Ther. 2008 Nov;7(11):3480-9
references.pmid: 18974392
references.citation: Pharmacology. 2001 May;62(4):208-12
references.pmid: 11359996
references.citation: Cancer Cell. 2008 Aug 12;14(2):111-22
references.pmid: 18656424
references.citation: Nat Rev Drug Discov. 2010 Oct;9(10):790-803
references.pmid: 20885410
references.citation: Cell Death Differ. 2012 Mar;19(3):369-77
references.pmid: 22223105
references.citation: Ann Surg Oncol. 2003 May;10(4):463-8
references.pmid: 12734097
references.citation: Int J Radiat Oncol Biol Phys. 1999 May 1;44(2):407-12
references.pmid: 10760437
references.citation: Nat Rev Cancer. 2008 Feb;8(2):121-32
references.pmid: 18202696
references.citation: BMC Cancer. 2008 Nov 03;8:318
references.pmid: 18980695
references.citation: Mol Cancer Ther. 2007 Jul;6(7):1920-31
references.pmid: 17620424
references.citation: Exp Cell Res. 1991 Jun;194(2):275-83
references.pmid: 2026179
references.citation: Mol Cell Biol. 2007 Jun;27(12):4513-25
references.pmid: 17438137
references.citation: Eur J Surg Oncol. 2001 Jun;27(4):340-2
references.pmid: 11417976
references.citation: N Engl J Med. 1995 Apr 13;332(15):1004-14
references.pmid: 7885406
references.citation: Ann Oncol. 2012 Mar;23(3):547-555
references.pmid: 21896541
references.citation: Cell Cycle. 2007 May 15;6(10):1200-4
references.pmid: 17471023
references.citation: J Cell Sci. 2003 Feb 1;116(Pt 3):513-24
references.pmid: 12508112
references.citation: Nat Cell Biol. 2008 Dec;10(12):1411-20
references.pmid: 18997788
references.citation: Cell Biol Int. 2000;24(2):61-70
references.pmid: 10772764
references.citation: Cell Death Differ. 2008 Jul;15(7):1153-62
references.pmid: 18404154
references.citation: Cancer. 2000 Jun 1;88(11):2619-28
references.pmid: 10861441
references.citation: Melanoma Res. 1998 Jun;8(3):240-4
references.pmid: 9664145
references.citation: Mol Interv. 2011 Apr;11(2):141-50
references.pmid: 21540474
references.citation: Nature. 2011 Mar 3;471(7336):104-9
references.pmid: 21368833
references.citation: Cancer Res. 1984 Oct;44(10 Suppl):4826s-4835s
references.pmid: 6467235
references.citation: Biochim Biophys Acta. 2008 Apr;1785(2):96-132
references.pmid: 18068131
references.citation: Oncologist. 2009 Aug;14(8):780-93
references.pmid: 19684075
references.citation: Mol Biol Cell. 2012 Jan;23(1):1-6
references.pmid: 22210845
references.citation: Eur J Pharm Biopharm. 2007 Jun;66(3):391-7
references.pmid: 17240125
references.citation: Cancer Cell. 2009 Oct 6;16(4):347-58
references.pmid: 19800579
references.citation: Oncogene. 2012 Jan 5;31(1):13-26
references.pmid: 21643015
references.citation: Biol Pharm Bull. 2013;36(3):376-83
references.pmid: 23229357
references.citation: Cancer Res. 2004 Apr 1;64(7):2502-8
references.pmid: 15059905
references.citation: Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):4106-11
references.pmid: 17360485