CAMK1D amplification implicated in epithelial–mesenchymal transition in basal‐like breast cancer

Breast cancer exhibits clinical and molecular heterogeneity, where expression profiling studies have identified five major molecular subtypes. The basal‐like subtype, expressing basal epithelial markers and negative for estrogen receptor (ER), progesterone receptor (PR) and HER2, is associated with higher overall levels of DNA copy number alteration (CNA), specific CNAs (like gain on chromosome 10p), and poor prognosis. Discovering the molecular genetic basis of tumor subtypes may provide new opportunities for therapy. To identify the driver oncogene on 10p associated with basal‐like tumors, we analyzed genomic profiles of 172 breast carcinomas. The smallest shared region of gain spanned just seven genes at 10p13, including calcium/calmodulin‐dependent protein kinase ID (CAMK1D), functioning in intracellular signaling but not previously linked to cancer. By microarray, CAMK1D was overexpressed when amplified, and by immunohistochemistry exhibited elevated expression in invasive carcinomas compared to carcinoma in situ. Engineered overexpression of CAMK1D in non‐tumorigenic breast epithelial cells led to increased cell proliferation, and molecular and phenotypic alterations indicative of epithelial–mesenchymal transition (EMT), including loss of cell–cell adhesions and increased cell migration and invasion. Our findings identify CAMK1D as a novel amplified oncogene linked to EMT in breast cancer, and as a potential therapeutic target with particular relevance to clinically unfavorable basal‐like tumors.

[1]  Ash A. Alizadeh,et al.  Genome-wide analysis of DNA copy-number changes using cDNA microarrays , 1999, Nature Genetics.

[2]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[3]  Marcel J. T. Reinders,et al.  SIRAC: Supervised Identification of Regions of Aberration in aCGH datasets , 2007, BMC Bioinformatics.

[4]  Dagny Faksvåg Haugen,et al.  Influence of TP 53 Gene Alterations and c-erbB2 Expression on the Response to Treatment with Doxorubicin in Locally Advanced Breast Cancer 1 , 2001 .

[5]  A. Shaywitz,et al.  CREB: a stimulus-induced transcription factor activated by a diverse array of extracellular signals. , 1999, Annual review of biochemistry.

[6]  Gavin Sherlock,et al.  The Stanford Microarray Database: implementation of new analysis tools and open source release of software , 2002, Nucleic Acids Res..

[7]  Hongjuan Zhao,et al.  TP53 mutation status and gene expression profiles are powerful prognostic markers of breast cancer , 2007, Breast Cancer Research.

[8]  Christian A. Rees,et al.  Molecular portraits of human breast tumours , 2000, Nature.

[9]  D. Tarin,et al.  Carcinoma invasion and metastasis: a role for epithelial-mesenchymal transition? , 2005, Cancer research.

[10]  A. Jemal,et al.  Cancer Statistics, 2007 , 2007, CA: a cancer journal for clinicians.

[11]  Charles M. Perou,et al.  A Comparison of Gene Expression Signatures from Breast Tumors and Breast Tissue Derived Cell Lines , 2002, Disease markers.

[12]  Tatiana A. Tatusova,et al.  NCBI Reference Sequence Project: update and current status , 2003, Nucleic Acids Res..

[13]  Robert Tibshirani,et al.  Distinct patterns of DNA copy number alteration are associated with different clinicopathological features and gene‐expression subtypes of breast cancer , 2006, Genes, chromosomes & cancer.

[14]  D. Huntsman,et al.  Tissue microarray analysis of neuroendocrine differentiation and its prognostic significance in breast cancer. , 2003, Human pathology.

[15]  T. Aas,et al.  Influence of TP53 gene alterations and c-erbB-2 expression on the response to treatment with doxorubicin in locally advanced breast cancer. , 2001, Cancer research.

[16]  M. Beato,et al.  Inducible regulatory elements in the human cyclin D1 promoter. , 1994, Oncogene.

[17]  Jill P. Mesirov,et al.  GSEA-P: a desktop application for Gene Set Enrichment Analysis , 2007, Bioinform..

[18]  Amy V Kapp,et al.  Discovery and validation of breast cancer subtypes , 2006, BMC Genomics.

[19]  Pablo Tamayo,et al.  Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[20]  J. Lammers,et al.  Characterization of the role of CaMKI-like kinase (CKLiK) in human granulocyte function. , 2005, Blood.

[21]  Jürgen Geisler,et al.  TP53 gene mutations predict the response to neoadjuvant treatment with 5-fluorouracil and mitomycin in locally advanced breast cancer. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[22]  A. Means,et al.  Regulation of Cyclin D1/Cdk4 Complexes by Calcium/Calmodulin-dependent Protein Kinase I* , 2004, Journal of Biological Chemistry.

[23]  A. Means,et al.  Ca(2+)/CaM-dependent kinases: from activation to function. , 2001, Annual review of pharmacology and toxicology.

[24]  J. Thiery.  Epithelial–mesenchymal transitions in tumour progression , 2002, Nature Reviews Cancer.

[25]  C. Sawyers.  Rational therapeutic intervention in cancer: kinases as drug targets. , 2002, Current opinion in genetics & development.

[26]  J. Lammers,et al.  Identification and characterization of CKLiK, a novel granulocyte Ca(++)/calmodulin-dependent kinase. , 2000, Blood.

[27]  G. Schuler.  Pieces of the puzzle: expressed sequence tags and the catalog of human genes , 1997, Journal of Molecular Medicine.

[28]  T. Soderling.  The Ca-calmodulin-dependent protein kinase cascade. , 1999, Trends in biochemical sciences.

[29]  David Botstein,et al.  Different gene expression patterns in invasive lobular and ductal carcinomas of the breast. , 2004, Molecular biology of the cell.

[30]  R. Tibshirani,et al.  Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[31]  C. Perou,et al.  Race, breast cancer subtypes, and survival in the Carolina Breast Cancer Study. , 2006, JAMA.

[32]  Christian A. Rees,et al.  Microarray analysis reveals a major direct role of DNA copy number alteration in the transcriptional program of human breast tumors , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[33]  W. Boecker,et al.  Evidence of progenitor cells of glandular and myoepithelial cell lineages in the human adult female breast epithelium: a new progenitor (adult stem) cell concept , 2003, Cell proliferation.

[34]  R. Tibshirani,et al.  Repeated observation of breast tumor subtypes in independent gene expression data sets , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[35]  Kenny Q. Ye,et al.  Novel patterns of genome rearrangement and their association with survival in breast cancer. , 2006, Genome research.

[36]  H. Tokumitsu,et al.  Identification and characterization of novel components of a Ca2+/calmodulin‐dependent protein kinase cascade in HeLa cells , 2003, FEBS letters.

[37]  R. Tibshirani,et al.  A method for calling gains and losses in array CGH data. , 2005, Biostatistics.

[38]  Harry Bartelink,et al.  Gene expression profiling and histopathological characterization of triple-negative/basal-like breast carcinomas , 2007, Breast Cancer Research.

[39]  N. Dubrawsky.  Cancer statistics , 1989, CA: a cancer journal for clinicians.

[40]  C. Isaacs,et al.  Utilizing prognostic and predictive factors in breast cancer , 2005, Current treatment options in oncology.

[41]  Ajay N. Jain,et al.  Genomic and transcriptional aberrations linked to breast cancer pathophysiologies. , 2006, Cancer cell.

[42]  J. McCubrey,et al.  Calcium/calmodulin-dependent kinase I and calcium/calmodulin-dependent kinase kinase participate in the control of cell cycle progression in MCF-7 human breast cancer cells. , 2005, Cancer research.