Abstract
To investigate the detailed molecular mechanism of mammary carcinogenesis and discover novel therapeutic targets, we previously analysed gene expression profiles of breast cancers. We here report characterization of a significant role of DTL/RAMP (denticleless/RA-regulated nuclear matrix associated protein) in mammary carcinogenesis. Semiquantitative RT–PCR and northern blot analyses confirmed upregulation of DTL/RAMP in the majority of breast cancer cases and all of breast cancer cell lines examined. Immunocytochemical and western blot analyses using anti-DTL/RAMP polyclonal antibody revealed cell-cycle-dependent localization of endogenous DTL/RAMP protein in breast cancer cells; nuclear localization was observed in cells at interphase and the protein was concentrated at the contractile ring in cytokinesis process. The expression level of DTL/RAMP protein became highest at G1/S phases, whereas its phosphorylation level was enhanced during mitotic phase. Treatment of breast cancer cells, T47D and HBC4, with small-interfering RNAs against DTL/RAMP effectively suppressed its expression and caused accumulation of G2/M cells, resulting in growth inhibition of cancer cells. We further demonstrate the in vitro phosphorylation of DTL/RAMP through an interaction with the mitotic kinase, Aurora kinase-B (AURKB). Interestingly, depletion of AURKB expression with siRNA in breast cancer cells reduced the phosphorylation of DTL/RAMP and decreased the stability of DTL/RAMP protein. These findings imply important roles of DTL/RAMP in growth of breast cancer cells and suggest that DTL/RAMP might be a promising molecular target for treatment of breast cancer.
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Adams RR, Carmena M, Earnshaw WC . (2001). Chromosomal passengers and the (aurora) ABCs of mitosis. Trends Cell Biol 11: 49–54.
Bange J, Zwick E, Ullrich A . (2001). Molecular targets for breast cancer therapy and prevention. Nat Med 7: 548–552.
Banks D, Wu M, Higa LA, Gavrilova N, Quan J, Ye T et al. (2006). L2DTL/CDT2 and PCNA interact with p53 and regulate p53 polyubiquitination and protein stability through MDM2 and CUL4A/DDB1 complexes. Cell Cycle 5: 1719–1729.
Carmena M, Earnshaw WC . (2003). The cellular geography of aurora kinases. Nat Rev Mol Cell Biol 4: 842–854.
Cheeseman IM, Anderson S, Jwa M, Green EM, Kang J, Yates III JR et al. (2002). Phospho-regulation of kinetochore-microtubule attachments by the Aurora kinase Ipl1p. Cell 111: 163–172.
Cheung WM, Chu AH, Chu PW, Ip NY . (2001). Cloning and expression of a novel nuclear matrix-associated protein that is regulated during the retinoic acid-induced neuronal differentiation. J Biol Chem 276: 17083–17091.
Dechat T, Gotzmann J, Stockinger A, Harris CA, Talle MA, Siekierka JJ et al. (1998). Detergent-salt resistance of LAP2alpha in interphase nuclei and phosphorylation-dependent association with chromosomes early in nuclear assembly implies functions in nuclear structure dynamics. EMBO J 17: 4887–4902.
Goto H, Yasui Y, Kawajiri A, Nigg EA, Terada Y, Tatsuka M et al. (2003). Aurora-B regulates the cleavage furrow-specific vimentin phosphorylation in the cytokinetic process. J Biol Chem 278: 8526–8530.
Higa LA, Banks D, Wu M, Kobayashi R, Sun H, Zhang H . (2006a). L2DTL/CDT2 interacts with the CUL4/DDB1 complex and PCNA and regulates CDT1 proteolysis in response to DNA damage. Cell Cycle 5: 1675–1680.
Higa LA, Wu M, Ye T, Kobayashi R, Sun H, Zhang H . (2006b). CUL4-DDB1 ubiquitin ligase interacts with multiple WD40-repeat proteins and regulates histone methylation. Nat Cell Biol 8: 1277–1283.
Hirota E, Yan L, Tsunoda T, Ashida S, Fujime M, Shuin T et al. (2006). Genome-wide gene expression profiles of clear cell renal cell carcinoma: identification of molecular targets for treatment of renal cell carcinoma. Int J Oncol 29: 799–827.
Lin ML, Park JH, Nishidate T, Nakamura Y, Katagiri T . (2007). Involvement of maternal embryonic leucine zipper kinase (MELK) in mammary carcinogenesis through interaction with Bcl-G, a pro-apoptotic member of Bcl-2 family. Breast Cancer Res 9: R17.
Nagayama S, Fukukawa C, Katagiri T, Okamoto T, Aoyama T, Oyaizu N et al. (2005). Therapeutic potential of antibodies against FZD 10, a cell-surface protein, for synovial sarcomas. Oncogene 24: 6201–6212.
Nagayama S, Iiizumi M, Katagiri T, Toguchida J, Nakamura Y . (2004). Identification of PDZK4, a novel human gene with PDZ domains, that is upregulated in synovial sarcomas. Oncogene 23: 5551–5557.
Navolanic PM, McCubrey JA . (2005). Pharmacological breast cancer therapy (review). Int J Oncol 27: 1341–1344.
Nishidate T, Katagiri T, Lin ML, Mano Y, Miki Y, Kasumi F et al. (2004). Genome-wide gene-expression profiles of breast-cancer cells purified with laser microbeam microdissection: identification of genes associated with progression and metastasis. Int J Oncol 25: 797–819.
Ohashi S, Sakashita G, Ban R, Nagasawa M, Matsuzaki H, Murata Y et al. (2006). Phospho-regulation of human protein kinase Aurora-A: analysis using anti-phospho-Thr288 monoclonal antibodies. Oncogene 25: 7691–7702.
Park JH, Lin ML, Nishidate T, Nakamura Y, Katagiri T . (2006). PDZ-binding kinase/T-LAK cell-originated protein kinase, a putative cancer/testis antigen having an oncogenic activity in breast cancer. Cancer Res 66: 9186–9195.
Parkin DM, Bray F, Ferlay J, Pisani P . (2005). Global cancer statistics, 2002. CA Cancer J Clin 55: 74–108.
Petricoin III EF, Hackett JL, Lesko LJ, Puri RK, Gutman SI, Chumakov K et al. (2002). Medical applications of microarray technologies: a regulatory science perspective. Nat Genet 32 (Suppl): 474–479.
Sansam CL, Shepard JL, Lai K, Ianari A, Danielian PS, Amsterdam A et al. (2006). DTL/CDT2 is essential for both CDT1 regulation and the early G2/M checkpoint. Genes Dev 20: 3117–3129.
Severson AF, Hamill DR, Carter JC, Schumacher J, Bowerman B . (2000). The aurora-related kinase AIR-2 recruits ZEN-4/CeMKLP1 to the mitotic spindle at metaphase and is required for cytokinesis. Curr Biol 10: 1162–1171.
Shimo A, Nishidate T, Ohta T, Fukuda M, Nakamura Y, Katagiri T . (2007). Elevated expression of PRC1, protein regulator of cytokinesis 1, involved in the growth of breast cancer cells. Cancer Sci 98: 174–181.
Shimokawa T, Furukawa Y, Sakai M, Li M, Miwa N, Lin YM et al. (2003). Involvement of the FGF18 gene in colorectal carcinogenesis, as a novel downstream target of the beta-catenin/T-cell factor complex. Cancer Res 63: 6116–6120.
Terada Y . (2001). Role of chromosomal passenger complex in chromosome segregation and cytokinesis. Cell Struct Funct 26: 653–657.
Togashi A, Katagiri T, Ashida S, Fujioka T, Maruyama O, Wakumoto Y et al. (2005). Hypoxia-inducible protein 2 (HIG2), a novel diagnostic marker for renal cell carcinoma and potential target for molecular therapy. Cancer Res 65: 4817–4826.
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We thank Ms Kyoko Kijima for excellent technical assistances.
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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)
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Ueki, T., Nishidate, T., Park, J. et al. Involvement of elevated expression of multiple cell-cycle regulator, DTL/RAMP (denticleless/RA-regulated nuclear matrix associated protein), in the growth of breast cancer cells. Oncogene 27, 5672–5683 (2008). https://doi.org/10.1038/onc.2008.186
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DOI: https://doi.org/10.1038/onc.2008.186
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