Progressive Telomere Dysfunction Causes Cytokinesis Failure and Leads to the Accumulation of Polyploid Cells
Most cancer cells accumulate genomic abnormalities at a remarkably rapid rate, as they are unable to maintain their chromosome structure and number. Excessively short telomeres, a known source of chromosome instability, are observed in early human-cancer lesions. Besides telomere dysfunction, it has been suggested that a transient phase of polyploidization, in most cases tetraploidization, has a causative role in cancer. Proliferation of tetraploids can gradually generate subtetraploid lineages of unstable cells that might fire the carcinogenic process by promoting further aneuploidy and genomic instability. Given the significance of telomere dysfunction and tetraploidy in the early stages of carcinogenesis, we investigated whether there is a connection between these two important promoters of chromosomal instability. We report that human mammary epithelial cells exhibiting progressive telomere dysfunction, in a pRb deficient and wild-type p53 background, fail to complete the cytoplasmatic cell division due to the persistence of chromatin bridges in the midzone. Flow cytometry together with fluorescence in situ hybridization demonstrated an accumulation of binucleated polyploid cells upon serial passaging cells. Restoration of telomere function through hTERT transduction, which lessens the formation of anaphase bridges by recapping the chromosome ends, rescued the polyploid phenotype. Live-cell imaging revealed that these polyploid cells emerged after abortive cytokinesis due to the persistence of anaphase bridges with large intervening chromatin in the cleavage plane. In agreement with a primary role of anaphase bridge intermediates in the polyploidization process, treatment of HMEC-hTERT cells with bleomycin, which produces chromatin bridges through illegimitate repair, resulted in tetraploid binucleated cells. Taken together, we demonstrate that human epithelial cells exhibiting physiological telomere dysfunction engender tetraploid cells through interference of anaphase bridges with the completion of cytokinesis. These observations shed light on the mechanisms operating during the initial stages of human carcinogenesis, as they provide a link between progressive telomere dysfunction and tetraploidy.
Vyšlo v časopise:
Progressive Telomere Dysfunction Causes Cytokinesis Failure and Leads to the Accumulation of Polyploid Cells. PLoS Genet 8(4): e32767. doi:10.1371/journal.pgen.1002679
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1002679
Souhrn
Most cancer cells accumulate genomic abnormalities at a remarkably rapid rate, as they are unable to maintain their chromosome structure and number. Excessively short telomeres, a known source of chromosome instability, are observed in early human-cancer lesions. Besides telomere dysfunction, it has been suggested that a transient phase of polyploidization, in most cases tetraploidization, has a causative role in cancer. Proliferation of tetraploids can gradually generate subtetraploid lineages of unstable cells that might fire the carcinogenic process by promoting further aneuploidy and genomic instability. Given the significance of telomere dysfunction and tetraploidy in the early stages of carcinogenesis, we investigated whether there is a connection between these two important promoters of chromosomal instability. We report that human mammary epithelial cells exhibiting progressive telomere dysfunction, in a pRb deficient and wild-type p53 background, fail to complete the cytoplasmatic cell division due to the persistence of chromatin bridges in the midzone. Flow cytometry together with fluorescence in situ hybridization demonstrated an accumulation of binucleated polyploid cells upon serial passaging cells. Restoration of telomere function through hTERT transduction, which lessens the formation of anaphase bridges by recapping the chromosome ends, rescued the polyploid phenotype. Live-cell imaging revealed that these polyploid cells emerged after abortive cytokinesis due to the persistence of anaphase bridges with large intervening chromatin in the cleavage plane. In agreement with a primary role of anaphase bridge intermediates in the polyploidization process, treatment of HMEC-hTERT cells with bleomycin, which produces chromatin bridges through illegimitate repair, resulted in tetraploid binucleated cells. Taken together, we demonstrate that human epithelial cells exhibiting physiological telomere dysfunction engender tetraploid cells through interference of anaphase bridges with the completion of cytokinesis. These observations shed light on the mechanisms operating during the initial stages of human carcinogenesis, as they provide a link between progressive telomere dysfunction and tetraploidy.
Zdroje
1. LengauerCKinzlerKVogelsteinB 1998 Genetic instabilities in human cancers. Nature 396 643 649
2. StorchovaZKufferC 2008 The consequences of tetraploidy and aneuploidy. J Cell Sci 121 3859 3866
3. OlaharskiASoteloRSolorza-LunaGGonsebattMGuzmanP 2006 Tetraploidy and chromosomal instability are early events during cervical carcinogenesis. Carcinogenesis 27 337 343
4. GalipeauPCowanDSanchezCBarrettMEmondM 1996 17p (p53) allelic losses, 4N (G2/tetraploid) populations, and progression to aneuploidy in Barrett's esophagus. Proc Natl Acad Sci U S A 93 7081 7084
5. MaleyC 2007 Multistage carcinogenesis in Barrett's esophagus. Cancer Lett 245 22 32
6. ShackneySSmithCMillerBBurholtDMurthaK 1989 Model for the genetic evolution of human solid tumors. Cancer Res 49 3344 3354
7. StorchovaZPellmanD 2004 From polyploidy to aneuploidy, genome instability and cancer. Nat Rev Mol Cell Biol 5 45 54
8. GanemNStorchovaZPellmanD 2007 Tetraploidy, aneuploidy and cancer. Curr Opin Genet Dev 17 157 162
9. MayerVAguileraA 1990 High levels of chromosome instability in polyploids of Saccharomyces cerevisiae. Mutat Res 231 177 186
10. FujiwaraTBandiMNittaMIvanovaEBronsonR 2005 Cytokinesis failure generating tetraploids promotes tumorigenesis in p53-null cells. Nature 437 1043 1047
11. StorchováZBrenemanACandeJDunnJBurbankK 2006 Genome-wide genetic analysis of polyploidy in yeast. Nature 443 541 547
12. NiggEA 2002 Centrosome aberrations: cause or consequence of cancer progression? Nature Rev Cancer 2 815 825
13. BoveriT 2008 Concerning the origin of malignant tumours by Theodor Boveri. Translated and annotated by Henry Harris. J Cell Sci 121 Suppl 1 1 84
14. MeekerAHicksJGabrielsonEStraussWDe MarzoA 2004 Telomere shortening occurs in subsets of normal breast epithelium as well as in situ and invasive carcinoma. Am J Pathol 164 925 935
15. ArtandiSEChangSLeeSLAlsonSGottliebGJ 2000 Telomere dysfunction promotes non-reciprocal translocations and epithelial cancers in mice. Nature 406 641 645
16. MeekerAKHicksJLPlatzEAMarchGEBennettCJ 2002 Telomere shortening is an early somatic DNA alteration in human prostate tumorigenesis. Cancer Res 62 6405 6409
17. MeekerAHicksJIacobuzio-DonahueCMontgomeryEWestraW 2004 Telomere length abnormalities occur early in the initiation of epithelial carcinogenesis. Clin Cancer Res 10 3317 3326
18. RomanovSKozakiewiczBHolstCStampferMHauptL 2001 Normal human mammary epithelial cells spontaneously escape senescence and acquire genomic changes. Nature 409 633 637
19. SolerDGenescàAArnedoGEgozcueJTusellL 2005 Telomere dysfunction drives chromosomal instability in human mammary epithelial cells. Genes Chromosomes Cancer 44 339 350
20. TusellLSolerDAgostiniMPampalonaJGenescàA 2008 The number of dysfunctional telomeres in a cell: one amplifies; more than one translocate. Cytogenet Genome Res 122 315 325
21. PampalonaJSolerDGenescàATusellL 2010 Whole chromosome loss is promoted by telomere dysfunction in primary cells. Genes Chromosomes Cancer 49 368 378
22. JuanGGongJTraganosFDarzynkiewiczZ 1996 Unscheduled expression of cyclins D1 and D3 in human tumour cell lines. Cell Prolif 29 259 266
23. StampferMGarbeJLevineGLichtsteinerSVasserotA 2001 Expression of the telomerase catalytic subunit, hTERT, induces resistance to transforming growth factor beta growth inhibition in p16INK4A(−) human mammary epithelial cells. Proc Natl Acad Sci U S A 98 4498 4503
24. MusaròMCiapponiLFasuloBGattiMCenciG 2008 Unprotected Drosophila melanogaster telomeres activate the spindle assembly checkpoint. Nat Genet 40 362 366
25. Lazzerini DenchiECelliGde LangeT 2006 Hepatocytes with extensive telomere deprotection and fusion remain viable and regenerate liver mass through endoreduplication. Genes Dev 20 2648 2653
26. HockemeyerDDanielsJTakaiHde LangeT 2006 Recent expansion of the telomeric complex in rodents: Two distinct POT1 proteins protect mouse telomeres. Cell 126 63 77
27. DavoliTDenchiEde LangeT 2010 Persistent telomere damage induces bypass of mitosis and tetraploidy. Cell 141 81 93
28. PanticMZimmermannSEl DalyHOpitzOGPoppS 2006 Telomere dysfunction and loss of p53 cooperate in defective mitotic segregation of chromosomes in cancer cells. Oncogene 25 4413 4420
29. MullinsJBieseleJ 1977 Terminal phase of cytokinesis in D-98s cells. J Cell Biol 73 672 684
30. ShiQKingRW 2005 Chromosome nondisjunction yields tetraploid rather than aneuploid cells in human cell lines. Nature 437 1038 1042
31. WeaverBASilkADClevelandDW 2006 Cell biology: nondisjunction, aneuploidy and tetraploidy. Nature 442 E9 10; discussion E10
32. SluderGNordbergJJ 2004 The good, the bad and the ugly: the practical consequences of centrosome amplification. Curr Opin Cell Biol 16 49 54
33. ShimizuNShingakiKKaneko-SasaguriYHashizumeTKandaT 2005 When, where and how the bridge breaks: anaphase bridge breakage plays a crucial role in gene amplification and HSR generation. Exp Cell Res 302 233 243
34. PampalonaJSolerDGenescàATusellL 2010 Telomere dysfunction and chromosome structure modulate the contribution of individual chromosomes in abnormal nuclear morphologies. Mutat Res 683 16 22
35. SteigemannPWurzenbergerCSchmitzMHeldMGuizettiJ 2009 Aurora B-mediated abscission checkpoint protects against tetraploidization. Cell 136 473 484
36. VagnarelliPHudsonDFRibeiroSATrinkle-MulcahyLSpenceJM 2006 Condensin and Repo-Man-PP1 co-operate in the regulation of chromosome architecture during mitosis. Nat Cell Biol 8 1133 1142
37. GietRGloverDM 2001 Drosophila aurora B kinase is required for histone H3 phosphorylation and condensin recruitment during chromosome condensation and to organize the central spindle during cytokinesis. J Cell Biol 152 669 682
38. SotilloRHernandoEDíaz-RodríguezETeruya-FeldsteinJCordón-CardoC 2007 Mad2 overexpression promotes aneuploidy and tumorigenesis in mice. Cancer Cell 11 9 23
39. ChangCJGouldingSEarnshawWCCarmenaM 2003 RNAi analysis reveals an unexpected role for topoisomerase II in chromosome arm congression to a metaphase plate. J Cell Sci 116 4715 4726
40. SchlegelBPStaritaLMParvinJD 2003 Overexpression of a protein fragment of RNA helicase A causes inhibition of endogenous BRCA1 function and defects in ploidy and cytokinesis in mammary epithelial cells. Oncogene 22 983 991
41. VinciguerraPGodinhoSAParmarKPellmanDD'AndreaAD 2010 Cytokinesis failure occurs in Fanconi anemia pathway-deficient murine and human bone marrow hematopoietic cells. J Clin Invest 120 3834 3842
42. LaulierCChengAStarkJM 2011 The relative efficiency of homology-directed repair has distinct effects on proper anaphase chromosome separation. Nucleic Acids Res 39 5935 5944
43. BrennerAJStampferMRAldazCM 1998 Increased p16 expression with first senescence arrest in human mammary epithelial cells and extended growth capacity with p16 inactivation. Oncogene 17 199 205
44. GenescàAPampalonaJFríasCDomínguezDTusellL 2011 Role of telomere dysfunction in genetic intratumor diversity. Adv Cancer Res 112 11 41
45. DavoliTde LangeT 2011 The causes and consequences of polyploidy in normal development and cancer. Annu Rev Cell Dev Biol 27 585 610
46. GorgoulisVGVassiliouLVKarakaidosPZacharatosPKotsinasA 2005 Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions. Nature 434 907 913
47. BartkovaJHorejsíZKoedKKrämerATortF 2005 DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis. Nature 434 864 870
48. GanemNGodinhoSPellmanD 2009 A mechanism linking extra centrosomes to chromosomal instability. Nature 460 278 282
49. SilkworthWNardiISchollLCiminiD 2009 Multipolar spindle pole coalescence is a major source of kinetochore mis-attachment and chromosome mis-segregation in cancer cells. PLoS ONE 4 e6564 doi:10.1371/journal.pone.0006564
50. MartínMGenescàALatreLRibasMMiróR 2003 Radiation-induced chromosome breaks in ataxia-telangiectasia cells remain open. Int J Radiat Biol 79 203 210
51. JuanGDarzynkiewiczZ 2001 Bivariate analysis of DNA content and expression of cyclin proteins. Curr Protoc Cytom Chapter 7 Unit 7.9
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Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
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