LMO 4 is an essential mediator of ErbB 2 / HER 2 / Neu-induced breast cancer cell cycle progression

ErbB2/HER2/Neu-overexpressing breast cancers are characterized by poor survival due to high proliferation and metastasis rates and identifying downstream targets of ErbB2 should facilitate developing novel therapies for this disease. Gene expression profiling revealed the transcriptional regulator LIM-only protein 4 [LMO4] is upregulated during ErbB2-induced mouse mammary gland tumorigenesis. While LMO4 is frequently overexpressed in breast cancer and LMO4overexpressing mice develop mammary epithelial tumors, the mechanisms involved are unknown. Herein, we report that LMO4 is a downstream target of ErbB2 and PI3K in ErbB2-dependent breast cancer cells. Furthermore, LMO4 silencing reduces proliferation of these cells, inducing a G2/M arrest that was associated with decreased cullin-3, an E3-ubiquitin ligase component important for mitosis. Loss of LMO4 subsequently results in reduced Cyclin D1 and Cyclin E. Further supporting a role for LMO4 in modulating proliferation by regulating cullin-3 expression, we found that LMO4 expression oscillates throughout the cell cycle with maximum expression occurring during G2/M and these changes precede oscillations in cullin-3 levels. LMO4 levels are also highest in high grade/less differentiated breast cancers, which are characteristically highly proliferative. We conclude that LMO4 is a novel cell cycle regulator with a key role in mediating ErbB2-induced proliferation, a hallmark of ErbB2-positive disease.

[1]  R. Kucherlapati,et al.  Genetic Mechanisms in Apc-Mediated Mammary Tumorigenesis , 2009, PLoS genetics.

[2]  Hae-Ahm Lee,et al.  Genistein-induced neuronal apoptosis and G2/M cell cycle arrest is associated with MDC1 up-regulation and PLK1 down-regulation. , 2007, European journal of pharmacology.

[3]  K. Lam,et al.  The LIM-only factor LMO4 regulates expression of the BMP7 gene through an HDAC2-dependent mechanism, and controls cell proliferation and apoptosis of mammary epithelial cells , 2007, Oncogene.

[4]  G. Sumara,et al.  A Cul3-based E3 ligase removes Aurora B from mitotic chromosomes, regulating mitotic progression and completion of cytokinesis in human cells. , 2007, Developmental cell.

[5]  J. Bergh,et al.  Strong Time Dependence of the 76-Gene Prognostic Signature for Node-Negative Breast Cancer Patients in the TRANSBIG Multicenter Independent Validation Series , 2007, Clinical Cancer Research.

[6]  Jin Xu,et al.  LMO4 mRNA stability is regulated by extracellular ATP in F11 cells. , 2007, Biochemical and biophysical research communications.

[7]  L. Prakash,et al.  ELA1 and CUL3 Are Required Along with ELC1 for RNA Polymerase II Polyubiquitylation and Degradation in DNA-Damaged Yeast Cells , 2007, Molecular and Cellular Biology.

[8]  Gregory E. J. Harding,et al.  Osteopontin localizes to the nucleus of 293 cells and associates with polo-like kinase-1. , 2007, American journal of physiology. Cell physiology.

[9]  Joshy George,et al.  Genetic reclassification of histologic grade delineates new clinical subtypes of breast cancer. , 2006, Cancer research.

[10]  Yusuke Nakamura,et al.  Gene expression profiles of small-cell lung cancers: molecular signatures of lung cancer. , 2006, International journal of oncology.

[11]  M. Dowsett,et al.  Trastuzumab after adjuvant chemotherapy in HER2-positive breast cancer. , 2005, The New England journal of medicine.

[12]  Greg Yothers,et al.  Trastuzumab plus adjuvant chemotherapy for operable HER2-positive breast cancer. , 2005, The New England journal of medicine.

[13]  Robert D Cardiff,et al.  The transcriptional repressor Snail promotes mammary tumor recurrence. , 2005, Cancer cell.

[14]  R. Keri,et al.  Gene expression profiling of cancer progression reveals intrinsic regulation of transforming growth factor-β signaling in ErbB2/Neu-induced tumors from transgenic mice , 2005, Oncogene.

[15]  J. Visvader,et al.  Loss of the LIM domain protein Lmo4 in the mammary gland during pregnancy impedes lobuloalveolar development , 2005, Oncogene.

[16]  J. Visvader,et al.  Overexpression of LMO4 induces mammary hyperplasia, promotes cell invasion, and is a predictor of poor outcome in breast cancer. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[17]  J. Visvader,et al.  The LIM Domain Protein Lmo4 Is Highly Expressed in Proliferating Mouse Epithelial Tissues , 2005, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[18]  E. Andrechek,et al.  Targeted disruption of ErbB2/Neu in the mammary epithelium results in impaired ductal outgrowth , 2005, Oncogene.

[19]  M. Gorospe,et al.  Concurrent versus individual binding of HuR and AUF1 to common labile target mRNAs , 2004, The EMBO journal.

[20]  B. Andersen,et al.  Expression of an engrailed-LMO4 fusion protein in mammary epithelial cells inhibits mammary gland development in mice , 2004, Oncogene.

[21]  J. Visvader,et al.  Mutational analysis of the LMO4 gene, encoding a BRCA1‐interacting protein, in breast carcinomas , 2003, International journal of cancer.

[22]  S. Elledge,et al.  BTB proteins are substrate-specific adaptors in an SCF-like modular ubiquitin ligase containing CUL-3 , 2003, Nature.

[23]  Sergio Wittlin,et al.  Two promoters within the human LMO4 gene contribute to its overexpression in breast cancer cells. , 2003, Genomics.

[24]  D. Stern,et al.  ErbBs in mammary development. , 2003, Experimental cell research.

[25]  C. Arteaga,et al.  Herceptin-induced inhibition of phosphatidylinositol-3 kinase and Akt Is required for antibody-mediated effects on p27, cyclin D1, and antitumor action. , 2002, Cancer research.

[26]  Richard Simon,et al.  Initiating oncogenic event determines gene-expression patterns of human breast cancer models , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[27]  Xin Yu,et al.  The LIM Domain Protein LMO4 Interacts with the Cofactor CtIP and the Tumor Suppressor BRCA1 and Inhibits BRCA1 Activity* , 2002, The Journal of Biological Chemistry.

[28]  Spyro Mousses,et al.  Clinical validation of candidate genes associated with prostate cancer progression in the CWR22 model system using tissue microarrays. , 2002, Cancer research.

[29]  C. Hudis,et al.  Cardiac dysfunction in the trastuzumab clinical trials experience. , 2002, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[30]  J. Visvader,et al.  The LIM domain gene LMO4 inhibits differentiation of mammary epithelial cells in vitro and is overexpressed in breast cancer , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[31]  A. Lenferink,et al.  ErbB2/neu kinase modulates cellular p27(Kip1) and cyclin D1 through multiple signaling pathways. , 2001, Cancer research.

[32]  Y. Geng,et al.  Specific protection against breast cancers by cyclin D1 ablation , 2001, Nature.

[33]  D. Watkins-Chow,et al.  Persistent Prop1 expression delays gonadotrope differentiation and enhances pituitary tumor susceptibility. , 2001, Human molecular genetics.

[34]  Myriam Gorospe,et al.  HuR regulates cyclin A and cyclin B1 mRNA stability during cell proliferation , 2000, The EMBO journal.

[35]  R. Keri,et al.  An NF-Y Binding Site Is Important for Basal, but Not Gonadotropin-releasing Hormone-stimulated, Expression of the Luteinizing Hormone β Subunit Gene* , 2000, The Journal of Biological Chemistry.

[36]  W. Muller,et al.  Signal transduction in mammary tumorigenesis: a transgenic perspective , 2000, Oncogene.

[37]  Y. Yarden,et al.  Cyclin D1 Is Required for Transformation by Activated Neu and Is Induced through an E2F-Dependent Signaling Pathway , 2000, Molecular and Cellular Biology.

[38]  M. Rosenfeld,et al.  Mouse deformed epidermal autoregulatory factor 1 recruits a LIM domain factor, LMO-4, and CLIM coregulators. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[39]  T. Rabbitts,et al.  LMO T-cell translocation oncogenes typify genes activated by chromosomal translocations that alter transcription and developmental processes. , 1998, Genes & development.

[40]  N. Hynes,et al.  ErbB‐2, the preferred heterodimerization partner of all ErbB receptors, is a mediator of lateral signaling , 1997, The EMBO journal.

[41]  M. Sliwkowski,et al.  Growth regulation of human breast and ovarian tumor cells by heregulin: Evidence for the requirement of ErbB2 as a critical component in mediating heregulin responsiveness. , 1996, Cancer research.

[42]  N. Hynes,et al.  Epidermal Growth Factor-related Peptides Activate Distinct Subsets of ErbB Receptors and Differ in Their Biological Activities (*) , 1996, The Journal of Biological Chemistry.

[43]  D. Caporossi,et al.  Characteristic chromosomal fragility of human embryonic cells exposed in vitro to aphidicolin , 1995, Human Genetics.

[44]  V. Sánchez-Margalet,et al.  Role of phosphatidylinositol-3-kinase in insulin receptor signaling: studies with inhibitor, LY294002. , 1994, Biochemical and biophysical research communications.

[45]  Y. Yarden,et al.  A single autophosphorylation site confers oncogenicity to the Neu/ErbB‐2 receptor and enables coupling to the MAP kinase pathway. , 1994, The EMBO journal.

[46]  M. Gossen,et al.  Acceleration of the G1/S phase transition by expression of cyclins D1 and E with an inducible system. , 1994, Molecular and cellular biology.

[47]  R. Cardiff,et al.  Expression of the neu protooncogene in the mammary epithelium of transgenic mice induces metastatic disease. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[48]  Y. Yarden,et al.  Regulated coupling of the Neu receptor to phosphatidylinositol 3'-kinase and its release by oncogenic activation. , 1992, The Journal of biological chemistry.

[49]  A. Ullrich,et al.  Oncogenic forms of the neu/HER2 tyrosine kinase are permanently coupled to phospholipase C gamma. , 1991, The EMBO journal.

[50]  Tony Hunter,et al.  Isolation of a human cyclin cDNA: Evidence for cyclin mRNA and protein regulation in the cell cycle and for interaction with p34cdc2 , 1989, Cell.

[51]  W Godolphin,et al.  Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. , 1989, Science.

[52]  M. Kraus,et al.  Overexpression of the EGF receptor‐related proto‐oncogene erbB‐2 in human mammary tumor cell lines by different molecular mechanisms. , 1987, The EMBO journal.

[53]  N. Nomura,et al.  Similarity of protein encoded by the human c-erb-B-2 gene to epidermal growth factor receptor , 1986, Nature.

[54]  J. Mosley,et al.  Cell cycle correlated genes dictate the prognostic power of breast cancer gene lists. , 2008, BMC medical genomics.

[55]  T. Barrette,et al.  ONCOMINE: a cancer microarray database and integrated data-mining platform. , 2004, Neoplasia.