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Breast Cancer Research
Published in: Breast Cancer Research 3/2010

Open Access 01-06-2010 | Research article

Senescence evasion by MCF-7 human breast tumor-initiating cells

Authors: Feridoun Karimi-Busheri, Aghdass Rasouli-Nia, John R Mackey, Michael Weinfeld

Published in: Breast Cancer Research | Issue 3/2010

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Abstract

Introduction

A subpopulation of cancer cells, tumor-initiating cells, is believed to be the driving force behind tumorigenesis and resistance to radiation and chemotherapy. The persistence of tumor-initiating cells may depend on altered regulation of DNA damage and checkpoint proteins, as well as a reduced propensity to undergo apoptosis or senescence.

Methods

To test this hypothesis, we isolated CD24-/low/CD44+ tumor-initiating cells (as mammospheres) from MCF-7 breast cancer cells grown in adherent monolayer culture, and carried out a comprehensive comparison of cell death and DNA damage response pathways prior to and after exposure to ionizing radiation in mammospheres and monolayer MCF-7 cells. Single and double-strand break repair was measured by single-cell gel electrophoresis. The latter was also examined by phosphorylation of histone H2AX and formation of 53BP1 and Rad51 foci. Apoptosis was quantified by flow-cytometric analysis of annexin V-binding and senescence was analyzed on the basis of cellular β-galactosidase activity. We employed the telomeric repeat amplification protocol to quantify telomerase activity. Expression of key DNA repair and cell cycle regulatory proteins was detected and quantified by western blot analysis.

Results

Our data demonstrate that in comparison to the bulk population of MCF-7 cells (predominantly CD24+/CD44+), the MCF-7 mammosphere cells benefit from a multifaceted approach to cellular protection relative to that seen in monolayer cells, including a reduced level of reactive oxygen species, a more active DNA single-strand break repair (SSBR) pathway, possibly due to a higher level of expression of the key SSBR protein, human AP endonuclease 1 (Ape1), and a significantly reduced propensity to undergo senescence as a result of increased telomerase activity and a low level of p21 protein expression. No significant difference was seen in the rates of double-strand break repair (DSBR) between the two cell types, but DSBR in mammospheres appears to by-pass the need for H2AX phosphorylation.

Conclusions

Enhanced survival of MCF-7 tumor-initiating cells in response to ionizing radiation is primarily dependent on an inherent down-regulation of the senescence pathway. Since MCF-7 cells are representative of cancer cells that do not readily undergo apoptosis, consideration of senescence pathways may play a role in targeting stem cells from such tumors.
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Literature
1.
Dumitrescu RG, Cotarla I: Understanding breast cancer risk -- where do we stand in 2005?. J Cell Mol Med. 2005, 9: 208-221. 10.1111/j.1582-4934.2005.tb00350.x.CrossRefPubMed
2.
Clarke MF, Fuller M: Stem cells and cancer: two faces of eve. Cell. 2006, 124: 1111-1115. 10.1016/j.cell.2006.03.011.CrossRefPubMed
3.
Ponti D, Zaffaroni N, Capelli C, Daidone MG: Breast cancer stem cells: an overview. Eur J Cancer. 2006, 42: 1219-1224. 10.1016/j.ejca.2006.01.031.CrossRefPubMed
4.
Eyler CE, Rich JN: Survival of the fittest: cancer stem cells in therapeutic resistance and angiogenesis. J Clin Oncol. 2008, 26: 2839-2845. 10.1200/JCO.2007.15.1829.CrossRefPubMedPubMedCentral
5.
Kai K, Arima Y, Kamiya T, Saya H: Breast cancer stem cells. Breast Cancer. 2010, 17: 80-85. 10.1007/s12282-009-0176-y.CrossRefPubMed
6.
Thomson CA, Giuliano AR, Shaw JW, Rock CL, Ritenbaugh CK, Hakim IA, Hollenbach KA, Alberts DS, Pierce JP: Diet and biomarkers of oxidative damage in women previously treated for breast cancer. Nutr Cancer. 2005, 51: 146-154. 10.1207/s15327914nc5102_4.CrossRefPubMed
7.
Kotsopoulos J, Narod SA: Towards a dietary prevention of hereditary breast cancer. Cancer Causes Control. 2005, 16: 125-138. 10.1007/s10552-004-2593-8.CrossRefPubMed
8.
Hoeijmakers JH: Genome maintenance mechanisms for preventing cancer. Nature. 2001, 411: 366-374. 10.1038/35077232.CrossRefPubMed
9.
Kronenwett R, Butterweck U, Steidl U, Kliszewski S, Neumann F, Bork S, Blanco ED, Roes N, Graf T, Brors B, Eils R, Maercker C, Kobbe G, Gattermann N, Haas R: Distinct molecular phenotype of malignant CD34(+) hematopoietic stem and progenitor cells in chronic myelogenous leukemia. Oncogene. 2005, 24: 5313-5324. 10.1038/sj.onc.1208596.CrossRefPubMed
10.
Bracker TU, Giebel B, Spanholtz J, Sorg UR, Klein-Hitpass L, Moritz T, Thomale J: Stringent regulation of DNA repair during human hematopoietic differentiation: a gene expression and functional analysis. Stem Cells. 2006, 24: 722-730. 10.1634/stemcells.2005-0227.CrossRefPubMed
11.
Chen MF, Lin CT, Chen WC, Yang CT, Chen CC, Liao SK, Liu JM, Lu CH, Lee KD: The sensitivity of human mesenchymal stem cells to ionizing radiation. Int J Radiat Oncol Biol Phys. 2006, 66: 244-253.CrossRefPubMed
12.
Van Sloun PP, Jansen JG, Weeda G, Mullenders LH, van Zeeland AA, Lohman PH, Vrieling H: The role of nucleotide excision repair in protecting embryonic stem cells from genotoxic effects of UV-induced DNA damage. Nucleic Acids Res. 1999, 27: 3276-3282. 10.1093/nar/27.16.3276.CrossRefPubMedPubMedCentral
13.
Saretzki G, Armstrong L, Leake A, Lako M, von Zglinicki T: Stress defense in murine embryonic stem cells is superior to that of various differentiated murine cells. Stem Cells. 2004, 22: 962-971. 10.1634/stemcells.22-6-962.CrossRefPubMed
14.
Hong Y, Stambrook PJ: Restoration of an absent G1 arrest and protection from apoptosis in embryonic stem cells after ionizing radiation. Proc Natl Acad Sci USA. 2004, 101: 14443-14448. 10.1073/pnas.0401346101.CrossRefPubMedPubMedCentral
15.
Cervantes RB, Stringer JR, Shao C, Tischfield JA, Stambrook PJ: Embryonic stem cells and somatic cells differ in mutation frequency and type. Proc Natl Acad Sci USA. 2002, 99: 3586-3590. 10.1073/pnas.062527199.CrossRefPubMedPubMedCentral
16.
Li X, Lewis MT, Huang J, Gutierrez C, Osborne CK, Wu MF, Hilsenbeck SG, Pavlick A, Zhang X, Chamness GC, Wong H, Rosen J, Chang JC: Intrinsic resistance of tumorigenic breast cancer cells to chemotherapy. J Natl Cancer Inst. 2008, 100: 672-679. 10.1093/jnci/djn123.CrossRefPubMed
17.
Pardal R, Clarke MF, Morrison SJ: Applying the principles of stem-cell biology to cancer. Nat Rev Cancer. 2003, 3: 895-902. 10.1038/nrc1232.CrossRefPubMed
18.
Fillmore CM, Kuperwasser C: Human breast cancer cell lines contain stem-like cells that self-renew, give rise to phenotypically diverse progeny and survive chemotherapy. Breast Cancer Res. 2008, 10: R25-10.1186/bcr1982.CrossRefPubMedPubMedCentral
19.
Charafe-Jauffret E, Ginestier C, Iovino F, Wicinski J, Cervera N, Finetti P, Hur MH, Diebel ME, Monville F, Dutcher J, Brown M, Viens P, Xerri L, Bertucci F, Stassi G, Dontu G, Birnbaum D, Wicha MS: Breast cancer cell lines contain functional cancer stem cells with metastatic capacity and a distinct molecular signature. Cancer Res. 2009, 69: 1302-1313. 10.1158/0008-5472.CAN-08-2741.CrossRefPubMedPubMedCentral
20.
Bao S, Wu Q, McLendon RE, Hao Y, Shi Q, Hjelmeland AB, Dewhirst MW, Bigner DD, Rich JN: Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature. 2006, 444: 756-760. 10.1038/nature05236.CrossRefPubMed
21.
Phillips TM, McBride WH, Pajonk F: The response of CD24(-/low)/CD44+ breast cancer-initiating cells to radiation. J Natl Cancer Inst. 2006, 98: 1777-1785. 10.1093/jnci/djj495.CrossRefPubMed
22.
Han JS, Crowe DL: Tumor initiating cancer stem cells from human breast cancer cell lines. Int J Oncol. 2009, 34: 1449-1453.PubMed
23.
Ponti D, Costa A, Zaffaroni N, Pratesi G, Petrangolini G, Coradini D, Pilotti S, Pierotti MA, Daidone MG: Isolation and in vitro propagation of tumorigenic breast cancer cells with stem/progenitor cell properties. Cancer Res. 2005, 65: 5506-5511. 10.1158/0008-5472.CAN-05-0626.CrossRefPubMed
24.
Collins AR: The comet assay for DNA damage and repair: principles, applications, and limitations. Mol Biotechnol. 2004, 26: 249-261. 10.1385/MB:26:3:249.CrossRefPubMed
25.
Lees-Miller SP, Godbout R, Chan DW, Weinfeld M, Day RS, Barron GM, Allalunis-Turner J: Absence of p350 subunit of DNA-activated protein kinase from a radiosensitive human cell line. Science. 1995, 267: 1183-1185. 10.1126/science.7855602.CrossRefPubMed
26.
Clayton H, Titley I, Vivanco M: Growth and differentiation of progenitor/stem cells derived from the human mammary gland. Exp Cell Res. 2004, 297: 444-460. 10.1016/j.yexcr.2004.03.029.CrossRefPubMed
27.
Goodell MA, McKinney-Freeman S, Camargo FD: Isolation and characterization of side population cells. Methods Mol Biol. 2005, 290: 343-352.PubMed
28.
Furuta T, Takemura H, Liao ZY, Aune GJ, Redon C, Sedelnikova OA, Pilch DR, Rogakou EP, Celeste A, Chen HT, Nussenzweig A, Aladjem MI, Bonner WM, Pommier Y: Phosphorylation of histone H2AX and activation of Mre11, Rad50, and Nbs1 in response to replication-dependent DNA double-strand breaks induced by mammalian DNA topoisomerase I cleavage complexes. J Biol Chem. 2003, 278: 20303-20312. 10.1074/jbc.M300198200.CrossRefPubMed
29.
Rasouli-Nia A, Karimi-Busheri F, Weinfeld M: Stable down-regulation of human polynucleotide kinase enhances spontaneous mutation frequency and sensitizes cells to genotoxic agents. Proc Natl Acad Sci USA. 2004, 101: 6905-6910. 10.1073/pnas.0400099101.CrossRefPubMedPubMedCentral
30.
Jardine LJ: Identification of senescence in cancer cells. Methods Mol Med. 2004, 88: 231-238.PubMed
31.
Parsons JL, Dianov GL: APE1 is the major 3'-phosphoglycolate activity in human cell extracts. Nucleic Acids Res. 2004, 32: 3531-3536. 10.1093/nar/gkh676.CrossRefPubMedPubMedCentral
32.
Izumi T, Brown DB, Naidu CV, Bhakat KK, Macinnes MA, Saito H, Chen DJ, Mitra S: Two essential but distinct functions of the mammalian abasic endonuclease. Proc Natl Acad Sci USA. 2005, 102: 5739-5743. 10.1073/pnas.0500986102.CrossRefPubMedPubMedCentral
33.
Fung H, Demple B: A vital role for Ape1/Ref1 protein in repairing spontaneous DNA damage in human cells. Mol Cell. 2005, 17: 463-470. 10.1016/j.molcel.2004.12.029.CrossRefPubMed
34.
Shrivastav M, De Haro LP, Nickoloff JA: Regulation of DNA double-strand break repair pathway choice. Cell Res. 2008, 18: 134-147. 10.1038/cr.2007.111.CrossRefPubMed
35.
Celeste A, Fernandez-Capetillo O, Kruhlak MJ, Pilch DR, Staudt DW, Lee A, Bonner RF, Bonner WM, Nussenzweig A: Histone H2AX phosphorylation is dispensable for the initial recognition of DNA breaks. Nat Cell Biol. 2003, 5: 675-679. 10.1038/ncb1004.CrossRefPubMed
36.
Xie A, Puget N, Shim I, Odate S, Jarzyna I, Bassing CH, Alt FW, Scully R: Control of sister chromatid recombination by histone H2AX. Mol Cell. 2004, 16: 1017-1025. 10.1016/j.molcel.2004.12.007.CrossRefPubMed
37.
Audebert M, Salles B, Calsou P: Involvement of poly(ADP-ribose) polymerase-1 and XRCC1/DNA ligase III in an alternative route for DNA double-strand breaks rejoining. J Biol Chem. 2004, 279: 55117-55126. 10.1074/jbc.M404524200.CrossRefPubMed
38.
Nakamura K, Sakai W, Kawamoto T, Bree RT, Lowndes NF, Takeda S, Taniguchi Y: Genetic dissection of vertebrate 53BP1: a major role in non-homologous end joining of DNA double strand breaks. DNA Repair (Amst). 2006, 5: 741-749. 10.1016/j.dnarep.2006.03.008.CrossRef
39.
Baumann P, West SC: Role of the human RAD51 protein in homologous recombination and double-stranded-break repair. Trends Biochem Sci. 1998, 23: 247-251. 10.1016/S0968-0004(98)01232-8.CrossRefPubMed
40.
Mukherjee B, Kessinger C, Kobayashi J, Chen BP, Chen DJ, Chatterjee A, Burma S: DNA-PK phosphorylates histone H2AX during apoptotic DNA fragmentation in mammalian cells. DNA Repair (Amst). 2006, 5: 575-590. 10.1016/j.dnarep.2006.01.011.CrossRef
41.
Essmann F, Engels IH, Totzke G, Schulze-Osthoff K, Janicke RU: Apoptosis resistance of MCF-7 breast carcinoma cells to ionizing radiation is independent of p53 and cell cycle control but caused by the lack of caspase-3 and a caffeine-inhibitable event. Cancer Res. 2004, 64: 7065-7072. 10.1158/0008-5472.CAN-04-1082.CrossRefPubMed
42.
Collado M, Gil J, Efeyan A, Guerra C, Schuhmacher AJ, Barradas M, Benguria A, Zaballos A, Flores JM, Barbacid M, Beach D, Serrano M: Tumour biology: senescence in premalignant tumours. Nature. 2005, 436: 642-10.1038/436642a.CrossRefPubMed
43.
Oh CW, Bump EA, Kim JS, Janigro D, Mayberg MR: Induction of a senescence-like phenotype in bovine aortic endothelial cells by ionizing radiation. Radiat Res. 2001, 156: 232-240. 10.1667/0033-7587(2001)156[0232:IOASLP]2.0.CO;2.CrossRefPubMed
44.
Elmore LW, Rehder CW, Di X, McChesney PA, Jackson-Cook CK, Gewirtz DA, Holt SE: Adriamycin-induced senescence in breast tumor cells involves functional p53 and telomere dysfunction. J Biol Chem. 2002, 277: 35509-35515. 10.1074/jbc.M205477200.CrossRefPubMed
45.
46.
Satra M, Tsougos I, Papanikolaou V, Theodorou K, Kappas C, Tsezou A: Correlation between radiation-induced telomerase activity and human telomerase reverse transcriptase mRNA expression in HeLa cells. Int J Radiat Biol. 2006, 82: 401-409. 10.1080/09553000600800090.CrossRefPubMed
47.
Bertram C, Hass R: Cellular responses to reactive oxygen species-induced DNA damage and aging. Biol Chem. 2008, 389: 211-220. 10.1515/BC.2008.031.CrossRefPubMed
48.
Zhang H: Molecular signaling and genetic pathways of senescence: Its role in tumorigenesis and aging. J Cell Physiol. 2007, 210: 567-574. 10.1002/jcp.20919.CrossRefPubMed
49.
Craig C, Kim M, Ohri E, Wersto R, Katayose D, Li Z, Choi YH, Mudahar B, Srivastava S, Seth P, Cowan K: Effects of adenovirus-mediated p16INK4A expression on cell cycle arrest are determined by endogenous p16 and Rb status in human cancer cells. Oncogene. 1998, 16: 265-272. 10.1038/sj.onc.1201493.CrossRefPubMed
50.
Russell M, Berardi P, Gong W, Riabowol K: Grow-ING, Age-ING and Die-ING: ING proteins link cancer, senescence and apoptosis. Exp Cell Res. 2006, 312: 951-961. 10.1016/j.yexcr.2006.01.020.CrossRefPubMed
51.
Nouman GS, Anderson JJ, Lunec J, Angus B: The role of the tumour suppressor p33 ING1b in human neoplasia. J Clin Pathol. 2003, 56: 491-496. 10.1136/jcp.56.7.491.CrossRefPubMedPubMedCentral
52.
Falck J, Mailand N, Syljuasen RG, Bartek J, Lukas J: The ATM-Chk2-Cdc25A checkpoint pathway guards against radioresistant DNA synthesis. Nature. 2001, 410: 842-847. 10.1038/35071124.CrossRefPubMed
53.
Stevens C, Smith L, La Thangue NB: Chk2 activates E2F-1 in response to DNA damage. Nat Cell Biol. 2003, 5: 401-409. 10.1038/ncb974.CrossRefPubMed
54.
Fujimoto H, Onishi N, Kato N, Takekawa M, Xu XZ, Kosugi A, Kondo T, Imamura M, Oishi I, Yoda A, Minami Y: Regulation of the antioncogenic Chk2 kinase by the oncogenic Wip1 phosphatase. Cell Death Differ. 2006, 13: 1170-1180. 10.1038/sj.cdd.4401801.CrossRefPubMed
55.
Hill R, Leidal AM, Madureira PA, Gillis LD, Cochrane HK, Waisman DM, Chiu A, Lee PW: Hypersensitivity to chromium-induced DNA damage correlates with constitutive deregulation of upstream p53 kinases in p21-/- HCT116 colon cancer cells. DNA Repair (Amst). 2008, 7: 239-252. 10.1016/j.dnarep.2007.10.001.CrossRef
56.
Minisini AM, Di Loreto C, Mansutti M, Artico D, Pizzolitto S, Piga A, Puglisi F: Topoisomerase IIalpha and APE/ref-1 are associated with pathologic response to primary anthracycline-based chemotherapy for breast cancer. Cancer Lett. 2005, 224: 133-139.CrossRefPubMed
57.
Lou Z, Chen J: Cellular senescence and DNA repair. Exp Cell Res. 2006, 312: 2641-2646. 10.1016/j.yexcr.2006.06.009.CrossRefPubMed
58.
Venot C, Maratrat M, Dureuil C, Conseiller E, Bracco L, Debussche L: The requirement for the p53 proline-rich functional domain for mediation of apoptosis is correlated with specific PIG3 gene transactivation and with transcriptional repression. EMBO J. 1998, 17: 4668-4679. 10.1093/emboj/17.16.4668.CrossRefPubMedPubMedCentral
59.
Baptiste N, Friedlander P, Chen X, Prives C: The proline-rich domain of p53 is required for cooperation with anti-neoplastic agents to promote apoptosis of tumor cells. Oncogene. 2002, 21: 9-21. 10.1038/sj.onc.1205015.CrossRefPubMed
Metadata
Title
Senescence evasion by MCF-7 human breast tumor-initiating cells
Authors
Feridoun Karimi-Busheri
Aghdass Rasouli-Nia
John R Mackey
Michael Weinfeld
Publication date
01-06-2010
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 3/2010
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/bcr2583

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