Differentiation of human embryonal carcinomas in vitro and in vivo reveals expression profiles relevant to normal development.

Embryonal carcinoma is a histologic subgroup of testicular germ cell tumors (TGCTs), and its cells may follow differentiation lineages in a manner similar to early embryogenesis. To acquire new knowledge about the transcriptional programs operating in this tumor development model, we used 22k oligo DNA microarrays to analyze normal and neoplastic tissue samples from human testis. Additionally, retinoic acid-induced in vitro differentiation was studied in relevant cell lines. We identified genes characterizing each of the known histologic subtypes, adding up to a total set of 687 differentially expressed genes. Among these, there was a significant overrepresentation of gene categories, such as genomic imprinting and gene transcripts associated to embryonic stem cells. Selection for genes highly expressed in the undifferentiated embryonal carcinomas resulted in the identification of 58 genes, including pluripotency markers, such as the homeobox genes NANOG and POU5F1 (OCT3/4), as well as GAL, DPPA4, and NALP7. Interestingly, abundant expression of several of the pluripotency genes was also detected in precursor lesions and seminomas. By use of tissue microarrays containing 510 clinical testicular samples, GAL and POU5F1 were up-regulated in TGCT also at the protein level and hence validated as diagnostic markers for undifferentiated tumor cells. The present study shows the unique gene expression profiles of each histologic subtype of TGCT from which we have identified deregulated components in selected processes operating in normal development, such as WNT signaling and DNA methylation.

[1]  R. Scully.  Special Tumors of Ovary and Testis , 1972 .

[2]  O. Dodge.  Histological Typing of Testis Tumours (International Histological Classification of Tumours, No. 16) , 1978, British Journal of Cancer.

[3]  P. Andrews,et al.  A comparative study of eight cell lines derived from human testicular teratocarcinoma , 1980, International journal of cancer.

[4]  P. Andrews,et al.  Retinoic acid fails to induce differentiation in human teratocarcinoma cell lines that express high levels of a cellular receptor protein. , 1983, Experimental cell research.

[5]  N C Dracopoli,et al.  Pluripotent embryonal carcinoma clones derived from the human teratocarcinoma cell line Tera-2. Differentiation in vivo and in vitro. , 1984, Laboratory investigation; a journal of technical methods and pathology.

[6]  P. Andrews.  Retinoic acid induces neuronal differentiation of a cloned human embryonal carcinoma cell line in vitro. , 1984, Developmental biology.

[7]  S. Lehnert,et al.  Temporal and regional changes in DNA methylation in the embryonic, extraembryonic and germ cell lineages during mouse embryo development. , 1987, Development.

[8]  P. Stern,et al.  Comparative analysis of cell surface antigens expressed by cell lines derived from human germ cell tumours , 1996, International journal of cancer.

[9]  A. Caricasole,et al.  Human growth-differentiation factor 3 (hGDF3): developmental regulation in human teratocarcinoma cell lines and expression in primary testicular germ cell tumours , 1998, Oncogene.

[10]  J. Thomson,et al.  Embryonic stem cell lines derived from human blastocysts. , 1998, Science.

[11]  H. Schöler,et al.  Formation of Pluripotent Stem Cells in the Mammalian Embryo Depends on the POU Transcription Factor Oct4 , 1998, Cell.

[12]  I. Sesterhenn,et al.  Histological Classification of Testis Tumours , 1998 .

[13]  C. Moskaluk,et al.  Activating c-kit gene mutations in human germ cell tumors. , 1999, The American journal of pathology.

[14]  R. Tibshirani,et al.  Significance analysis of microarrays applied to the ionizing radiation response , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[15]  Jason E. Stewart,et al.  Minimum information about a microarray experiment (MIAME)—toward standards for microarray data , 2001, Nature Genetics.

[16]  May D. Wang,et al.  GoMiner: a resource for biological interpretation of genomic and proteomic data , 2003, Genome Biology.

[17]  R. Lothe,et al.  New insights into testicular germ cell tumorigenesis from gene expression profiling. , 2002, Cancer research.

[18]  R. Kucherlapati,et al.  Genomic and expression analysis of the 12p11-p12 amplicon using EST arrays identifies two novel amplified and overexpressed genes. , 2002, Cancer research.

[19]  J. Thomson,et al.  BMP4 initiates human embryonic stem cell differentiation to trophoblast , 2002, Nature Biotechnology.

[20]  Abraham L. Kierszenbaum.  Genomic imprinting and epigenetic reprogramming: Unearthing the garden of forking paths , 2002, Molecular reproduction and development.

[21]  J. Thomson,et al.  Preimplantation Human Embryos and Embryonic Stem Cells Show Comparable Expression of Stage‐Specific Embryonic Antigens , 2002, Stem cells.

[22]  P. Andrews,et al.  From teratocarcinomas to embryonic stem cells. , 2002, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[23]  R. Lothe,et al.  Candidate genes for testicular cancer evaluated by in situ protein expression analyses on tissue microarrays. , 2003, Neoplasia.

[24]  S. Fosså,et al.  Stem cell factor receptor (c-KIT) codon 816 mutations predict development of bilateral testicular germ-cell tumors. , 2003, Cancer research.

[25]  C. Dennis.  Developmental reprogramming: Take a cell, any cell... , 2003, Nature.

[26]  M. Murakami,et al.  The Homeoprotein Nanog Is Required for Maintenance of Pluripotency in Mouse Epiblast and ES Cells , 2003, Cell.

[27]  J. Nichols,et al.  Functional Expression Cloning of Nanog, a Pluripotency Sustaining Factor in Embryonic Stem Cells , 2003, Cell.

[28]  P. Brown,et al.  Gene expression patterns in human embryonic stem cells and human pluripotent germ cell tumors , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[29]  Yusuke Nakamura,et al.  Analysis of gene-expression profiles in testicular seminomas using a genome-wide cDNA microarray. , 2003, International journal of oncology.

[30]  R. Huddart,et al.  Expression profile of genes from 12p in testicular germ cell tumors of adolescents and adults associated with i(12p) and amplification at 12p11.2–p12.1 , 2003, Oncogene.

[31]  Yusuke Nakamura,et al.  Oncogenic role of NALP7 in testicular seminomas , 2004, Cancer science.

[32]  Christian Schwager,et al.  Embryonic Stem Cell-Like Features of Testicular Carcinoma in Situ Revealed by Genome-Wide Gene Expression Profiling , 2004, Cancer Research.

[33]  A. Clark,et al.  Human STELLAR, NANOG, and GDF3 Genes Are Expressed in Pluripotent Cells and Map to Chromosome 12p13, a Hotspot for Teratocarcinoma , 2004, Stem cells.

[34]  O. Cummings,et al.  KIT mutations are common in testicular seminomas. , 2004, The American journal of pathology.

[35]  R. Puri,et al.  Gene expression in human embryonic stem cell lines: unique molecular signature. , 2004, Blood.