Abstract
Many types of cancer display heterogeneity in various features, including gene expression and malignant potential. This heterogeneity is associated with drug resistance and cancer progression. Recent studies have shown that the expression of a major protein quality control ubiquitin ligase, carboxyl terminus of Hsc70-interacting protein (CHIP), is negatively correlated with breast cancer clinicopathological stages and poor overall survival. Here we show that CHIP acts as a capacitor of heterogeneous Bcl-2 expression levels and prevents an increase in the anticancer drug-resistant population in breast cancer cells. CHIP knockdown in breast cancer cells increased variation in Bcl-2 expression levels, an antiapoptotic protein, among the cells. Our results also showed that CHIP knockdown increased the proportion of anticancer drug-resistant cells. These findings suggest that CHIP buffers variation in gene expression levels, affecting resistance to anticancer drugs. In single-cell clones derived from breast cancer cell lines, CHIP knockdown did not alter the variation in Bcl-2 expression levels and the proportion of anticancer drug-resistant cells. In contrast, when clonal cells were treated with a mutagen, the variation in Bcl-2 expression levels and proportion of anticancer drug-resistant cells were altered by CHIP knockdown. These results suggest that CHIP masks genetic variations to suppress heterogeneous Bcl-2 expression levels and prevents augmentation of the anticancer drug-resistant population of breast cancer cells. Because genetic variation is a major driver of heterogeneity, our results suggest that the degree of heterogeneity in expression levels is decided by a balance between genetic variation and the buffering capacity of CHIP.
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References
Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM . Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 2010; 127: 2893–2917.
Polyak K . Breast cancer: origins and evolution. J Clin Invest 2007; 117: 3155–3163.
Heng HH, Bremer SW, Stevens JB, Ye KJ, Liu G, Ye CJ . Genetic and epigenetic heterogeneity in cancer: a genome-centric perspective. J Cell Physiol 2009; 220: 538–547.
Lambert G, Estevez-Salmeron L, Oh S, Liao D, Emerson BM, Tlsty TD et al. An analogy between the evolution of drug resistance in bacterial communities and malignant tissues. Nat Rev Cancer 2011; 11: 375–382.
Yang JY, Ha SA, Yang YS, Kim JW . p-Glycoprotein ABCB5 and YB-1 expression plays a role in increased heterogeneity of breast cancer cells: correlations with cell fusion and doxorubicin resistance. BMC cancer 2010; 10: 388.
Ding L, Ley TJ, Larson DE, Miller CA, Koboldt DC, Welch JS et al. Clonal evolution in relapsed acute myeloid leukaemia revealed by whole-genome sequencing. Nature 2012; 481: 506–510.
Longley DB, Johnston PG . Molecular mechanisms of drug resistance. J Pathol 2005; 205: 275–292.
Coley HM . Mechanisms and consequences of chemotherapy resistance in breast cancer. Ejc Suppl 2009; 7: 3–7.
Rudin CM, Yang Z, Schumaker LM, VanderWeele DJ, Newkirk K, Egorin MJ et al. Inhibition of glutathione synthesis reverses Bcl-2-mediated cisplatin resistance. Cancer Res 2003; 63: 312–318.
Davis JM, Navolanic PM, Weinstein-Oppenheimer CR, Steelman LS, Hu W, Konopleva M et al. Raf-1 and Bcl-2 induce distinct and common pathways that contribute to breast cancer drug resistance. Clin Cancer Res 2003; 9: 1161–1170.
Lima RT, Martins LM, Guimaraes JE, Sambade C, Vasconcelos MH . Specific downregulation of bcl-2 and xIAP by RNAi enhances the effects of chemotherapeutic agents in MCF-7 human breast cancer cells. Cancer Gene Ther 2004; 11: 309–316.
Tabuchi Y, Matsuoka J, Gunduz M, Imada T, Ono R, Ito M et al. Resistance to paclitaxel therapy is related with Bcl-2 expression through an estrogen receptor mediated pathway in breast cancer. Int J Oncol 2009; 34: 313–319.
Tsujimoto Y, Shimizu S . VDAC regulation by the Bcl-2 family of proteins. Cell Death Differ 2000; 7: 1174–1181.
Youle RJ, Strasser A . The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 2008; 9: 47–59.
Sierra A, Castellsague X, Tortola S, Escobedo A, Lloveras B, Peinado MA et al. Apoptosis loss and bcl-2 expression: key determinants of lymph node metastases in T1 breast cancer. Clin Cancer Res 1996; 2: 1887–1894.
Kajiro M, Hirota R, Nakajima Y, Kawanowa K, So-ma K, Ito I et al. The ubiquitin ligase CHIP acts as an upstream regulator of oncogenic pathways. Nat Cell Biol 2009; 11: 312–319.
Ballinger CA, Connell P, Wu Y, Hu Z, Thompson LJ, Yin LY et al. Identification of CHIP, a novel tetratricopeptide repeat-containing protein that interacts with heat shock proteins and negatively regulates chaperone functions. Mol Cell Biol 1999; 19: 4535–4545.
Jiang J, Ballinger CA, Wu Y, Dai Q, Cyr DM, Hohfeld J et al. CHIP is a U-box-dependent E3 ubiquitin ligase: identification of Hsc70 as a target for ubiquitylation. J Biol Chem 2001; 276: 42938–42944.
Murata S, Minami Y, Minami M, Chiba T, Tanaka K . CHIP is a chaperone-dependent E3 ligase that ubiquitylates unfolded protein. EMBO Rep 2001; 2: 1133–1138.
Goldberg AL . Protein degradation and protection against misfolded or damaged proteins. Nature 2003; 426: 895–899.
Patani N, Jiang W, Newbold R, Mokbel K . Prognostic implications of carboxyl-terminus of Hsc70 interacting protein and lysyl-oxidase expression in human breast cancer. J Carcinog 2010; 9: 9.
Drewinko B, Brown BW, Gottlieb JA . The effect of cis-diamminedichloroplatinum (II) on cultured human lymphoma cells and its therapeutic implications. Cancer Res 1973; 33: 3091–3095.
Kampinga HH, Kanon B, Salomons FA, Kabakov AE, Patterson C . Overexpression of the cochaperone CHIP enhances Hsp70-dependent folding activity in mammalian cells. Mol Cell Biol 2003; 23: 4948–4958.
Rutherford SL, Lindquist S . Hsp90 as a capacitor for morphological evolution. Nature 1998; 396: 336–342.
Queitsch C, Sangster TA, Lindquist S . Hsp90 as a capacitor of phenotypic variation. Nature 2002; 417: 618–624.
Sollars V, Lu X, Xiao L, Wang X, Garfinkel MD, Ruden DM . Evidence for an epigenetic mechanism by which Hsp90 acts as a capacitor for morphological evolution. Nat Genet 2003; 33: 70–74.
Papendorp JT, Schatz RW, Soto AM, Sonnenschein C . On the role of 17 alpha-estradiol and 17 beta-estradiol in the proliferation of MCF7 and T47D-A11 human breast tumor cells. J Cell Physiol 1985; 125: 591–595.
Butler WB, Berlinski PJ, Hillman RM, Kelsey WH, Toenniges MM . Relation of in vitro properties to tumorigenicity for a series of sublines of the human breast cancer cell line MCF-7. Cancer Res 1986; 46: 6339–6348.
Resnicoff M, Medrano EE, Podhajcer OL, Bravo AI, Bover L, Mordoh J . Subpopulations of MCF7 cells separated by Percoll gradient centrifugation: a model to analyze the heterogeneity of human breast cancer. Proc Natl Acad Sci USA 1987; 84: 7295–7299.
McDiarmid HM, Douglas GR, Coomber BL, Josephy PD . Epithelial and fibroblast cell lines cultured from the transgenic BigBlue rat: an in vitro mutagenesis assay. Mutat Res 2001; 497: 39–47.
Young RL, Korsmeyer SJ . A negative regulatory element in the bcl-2 5′-untranslated region inhibits expression from an upstream promoter. Mol Cell Biol 1993; 13: 3686–3697.
Lee JH, Jeon MH, Seo YJ, Lee YJ, Ko JH, Tsujimoto Y et al. CA repeats in the 3′-untranslated region of bcl-2 mRNA mediate constitutive decay of bcl-2 mRNA. J Biol Chem 2004; 279: 42758–42764.
Miyashita T, Harigai M, Hanada M, Reed JC . Identification of a p53-dependent negative response element in the bcl-2 gene. Cancer Res 1994; 54: 3131–3135.
Hewitt SM, Hamada S, McDonnell TJ, Rauscher FJ 3rd, Saunders GF . Regulation of the proto-oncogenes bcl-2 and c-myc by the Wilms' tumor suppressor gene WT1. Cancer Res 1995; 55: 5386–5389.
Wilson BE, Mochon E, Boxer LM . Induction of bcl-2 expression by phosphorylated CREB proteins during B-cell activation and rescue from apoptosis. Mol Cell Biol 1996; 16: 5546–5556.
Kurland JF, Kodym R, Story MD, Spurgers KB, Mcdonnell TJ, Meyn RE . NF-kappaB1 (p50) homodimers contribute to transcription of the bcl-2 oncogene. J Biol Chem 2001; 276: 45380–45386.
Gomez-Manzano C, Mitlinga P, Fueyo J, Lee HY, Hu M, Spurgers KB et al. Transfer of E2F-1 to human glioma cells results in transcriptional up-regulation of Bcl-2. Cancer Res 2001; 61: 6693–6697.
Decary S, Decesse JT, Ogryzko V, Reed JC, Naguibneva I, Harel-Bellan A et al. The retinoblastoma protein binds the promoter of the survival gene bcl-2 and regulates its transcription in epithelial cells through transcription factor AP-2. Mol Cell Biol 2002; 22: 7877–7888.
Stevenson HS, Fu SW, Pinzone JJ, Rheey J, Simmens SJ, Berg PE . BP1 transcriptionally activates bcl-2 and inhibits TNFalpha-induced cell death in MCF7 breast cancer cells. Breast Cancer Res 2007; 9: R60.
Banerji S, Cibulskis K, Rangel-Escareno C, Brown KK, Carter SL, Frederick AM et al. Sequence analysis of mutations and translocations across breast cancer subtypes. Nature 2012; 486: 405–409.
Curtis C, Shah SP, Chin SF, Turashvili G, Rueda OM, Dunning MJ et al. The genomic and transcriptomic architecture of 2,000 breast tumours reveals novel subgroups. Nature 2012; 486: 346–352.
Magee JA, Piskounova E, Morrison SJ . Cancer stem cells: impact, heterogeneity, and uncertainty. Cancer Cell 2012; 21: 283–296.
Marusyk A, Almendro V, Polyak K . Intra-tumour heterogeneity: a looking glass for cancer? Nat Rev Cancer 2012; 12: 323–334.
Theodoraki MA, Caplan AJ . Quality control and fate determination of Hsp90 client proteins. Biochim Biophys Acta 2012; 1823: 683–688.
Acknowledgements
This work was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan. We would like to thank Enago (www.enago.jp) for the English language review.
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Tsuchiya, M., Nakajima, Y., Waku, T. et al. CHIP buffers heterogeneous Bcl-2 expression levels to prevent augmentation of anticancer drug-resistant cell population. Oncogene 34, 4656–4663 (2015). https://doi.org/10.1038/onc.2014.387
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DOI: https://doi.org/10.1038/onc.2014.387
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