Identification of molecular markers and signaling pathway in endometrial cancer in Hong Kong Chinese women by genome-wide gene expression profiling

Endometrial cancer is the third most common gynecologic malignancy and the ninth most common malignancy for females overall in Hong Kong. Approximately 80% or more of these cancers are endometrioid endometrial adenocarcinomas. The aim of this study was to reveal genes contributing to the development of endometrioid endometrial cancer, which may impact diagnosis, prognosis and treatment of the disease. Whole-genome gene expression analysis was completed for a set of 55 microdissected sporadic endometrioid endometrial adenocarcinomas and 29 microdissected normal endometrium specimens using the Affymetrix Human U133 Plus 2.0 oligonucleotide microarray. Selected genes of interest were validated by quantitative real-time-polymerase chain reaction (qRT-PCR). Pathway analysis was performed to reveal gene interactions involved in endometrial tumorigenesis. Unsupervised hierarchical clustering displayed a distinct separation between the endometrioid adenocarcinomas and normal endometrium samples. Supervised analysis identified 117 highly differentially regulated genes (⩾4.0-fold change), which distinguished the endometrial cancer specimens from normal endometrium. Twelve novel genes including DKK4, ZIC1, KIF1A, SAA2, LOC16378, ALPP2, CCL20, CXCL5, BST2, OLFM1, KLRC1 and MBC45780 were deregulated in the endometrial cancer, and further validated in an independent set of 56 cancer and 29 normal samples using qRT-PCR. In addition, 10 genes were differentially regulated in late-stage cancer, as compared to early-stage disease, and may be involved in tumor progression. Pathway analysis of the expression data from this tumor revealed an interconnected network consisting of 21 aberrantly regulated genes involved in angiogenesis, cell proliferation and chromosomal instability. The results of this study highlight the molecular features of endometrioid endometrial cancer and provide insight into the events underlying the development and progression of endometrioid endometrial cancer.

[1]  R. Kempson,et al.  Uterine papillary serous carcinoma: A highly malignant form of endometrial adenocarcinoma , 1982, The American journal of surgical pathology.

[2]  S. Sukumar,et al.  Cyclin E Is a Target of WT1 Transcriptional Repression* , 2002, The Journal of Biological Chemistry.

[3]  R. Reeves,et al.  Architectural Transcription Factor HMGI(Y) Promotes Tumor Progression and Mesenchymal Transition of Human Epithelial Cells , 2001, Molecular and Cellular Biology.

[4]  Ju-Hyung Woo,et al.  Increased Expression of Mitotic Checkpoint Genes in Breast Cancer Cells with Chromosomal Instability , 2006, Clinical Cancer Research.

[5]  A. Berchuck,et al.  Gene expression profiling of microsatellite unstable and microsatellite stable endometrial cancers indicates distinct pathways of aberrant signaling. , 2005, Cancer research.

[6]  J. Van Damme,et al.  The CC chemokine CCL20 and its receptor CCR6. , 2003, Cytokine & growth factor reviews.

[7]  R. Ross,et al.  Inhibition of PDGF‐stimulated and matrix‐mediated proliferation of human vascular smooth muscle cells by SPARC is independent of changes in cell shape or cyclin‐dependent kinase inhibitors , 2002, Journal of cellular biochemistry.

[8]  Li Yan,et al.  The Matrix Metalloproteinase-9/Neutrophil Gelatinase-Associated Lipocalin Complex Plays a Role in Breast Tumor Growth and Is Present in the Urine of Breast Cancer Patients , 2005, Clinical Cancer Research.

[9]  H. Koeffler,et al.  c-myb transactivates the human cyclin A1 promoter and induces cyclin A1 gene expression. , 1999, Blood.

[10]  Paul J. L. Zhang,et al.  WT1 is Differentially Expressed in Serous, Endometrioid, Clear Cell, and Mucinous Carcinomas of the Peritoneum, Fallopian Tube, Ovary, and Endometrium , 2004, International journal of gynecological pathology : official journal of the International Society of Gynecological Pathologists.

[11]  R. Gentleman,et al.  Global expression changes of constitutive and hormonally regulated genes during endometrial neoplastic transformation. , 2001, Gynecologic oncology.

[12]  M. Fini,et al.  Transcription Factors Pax6 and AP-2α Interact To Coordinate Corneal Epithelial Repair by Controlling Expression of Matrix Metalloproteinase Gelatinase B , 2004, Molecular and Cellular Biology.

[13]  J. Cigudosa,et al.  Differential gene expression profile in endometrioid and nonendometrioid endometrial carcinoma: STK15 is frequently overexpressed and amplified in nonendometrioid carcinomas. , 2003, Cancer research.

[14]  M. Detmar,et al.  Thrombospondin-2: a potent endogenous inhibitor of tumor growth and angiogenesis. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[15]  A. Kaneko,et al.  Endorphin and enkephalin ameliorate excessive synovial cell functions in patients with rheumatoid arthritis. , 2001, The Journal of rheumatology.

[16]  G. Murphy,et al.  Relating matrix metalloproteinase structure to function: why the "hemopexin" domain? , 1997, Matrix biology : journal of the International Society for Matrix Biology.

[17]  Hiroshi Kijima,et al.  Overexpression of the thrombospondin 2 (TSP2) gene modulated by the matrix metalloproteinase family expression and production in human colon carcinoma cell line. , 2003, Oncology reports.

[18]  C. Burger,et al.  Gene expression profiling in human endometrial cancer tissue samples: utility and diagnostic value. , 2004, Gynecologic oncology.

[19]  N. Socci,et al.  Distinctive Gene Expression Profiles by cDNA Microarrays in Endometrioid and Serous Carcinomas of the Endometrium , 2004, International journal of gynecological pathology : official journal of the International Society of Gynecological Pathologists.

[20]  M. Kitagawa,et al.  Activation of Cyclin-dependent Kinase 2 (Cdk2) in Growth-stimulated Rat Astrocytes , 1998, The Journal of Biological Chemistry.

[21]  Cheng Li,et al.  DNA-Chip Analyzer (dChip) , 2003 .

[22]  K. Miller,et al.  Cyclin A1 is highly expressed in aggressive testicular germ cell tumors. , 2003, Cancer letters.

[23]  R. Lupu,et al.  HMG-I(Y) recognizes base-unpairing regions of matrix attachment sequences and its increased expression is directly linked to metastatic breast cancer phenotype. , 1999, Cancer research.

[24]  A. Harris,et al.  Expression and hormone regulation of Wnt2, 3, 4, 5a, 7a, 7b and 10b in normal human endometrium and endometrial carcinoma. , 1997, British Journal of Cancer.

[25]  R. C. Johnston,et al.  Distinct chromosome segregation roles for spindle checkpoint proteins. , 2002, Molecular biology of the cell.

[26]  V. Abeler,et al.  Clear cell carcinoma of the endometrium: a histopathological and clinical study of 97 cases. , 1990, Gynecologic oncology.

[27]  N. Suzumori,et al.  Peritoneal fluid concentrations of epithelial neutrophil-activating peptide-78 correlate with the severity of endometriosis. , 2004, Fertility and sterility.

[28]  Wing H Wong,et al.  Expression profiling of serous low malignant potential, low-grade, and high-grade tumors of the ovary. , 2005, Cancer research.

[29]  K. Zatloukal,et al.  Inverse correlation of secreted frizzled‐related protein 4 and β‐catenin expression in endometrial stromal sarcomas , 2004, The Journal of pathology.

[30]  C. Teng,et al.  Characterization of estrogen-responsive mouse lactoferrin promoter. , 1991, The Journal of biological chemistry.

[31]  Yudong D. He,et al.  A cell proliferation signature is a marker of extremely poor outcome in a subpopulation of breast cancer patients. , 2005, Cancer research.

[32]  Kathleen R. Cho,et al.  p53 gene mutations are common in uterine serous carcinoma and occur early in their pathogenesis. , 1997, The American journal of pathology.

[33]  M. Simons,et al.  Thrombospondin Type 1 Repeats Interact with Matrix Metalloproteinase 2 , 2000, The Journal of Biological Chemistry.

[34]  P. Lehtovirta,et al.  Suppressed expression of insulin‐like growth factor binding protein‐1 mRNA in the endometrium: A molecular mechanism associating endometrial cancer with its risk factors , 1994, International journal of cancer.

[35]  Li Yan,et al.  The High Molecular Weight Urinary Matrix Metalloproteinase (MMP) Activity Is a Complex of Gelatinase B/MMP-9 and Neutrophil Gelatinase-associated Lipocalin (NGAL) , 2001, The Journal of Biological Chemistry.

[36]  W. Stetler-Stevenson,et al.  TIMP-2: an endogenous inhibitor of angiogenesis. , 2005, Trends in molecular medicine.

[37]  R. Khokha,et al.  Matrix metalloproteinases and their tissue inhibitors direct cell fate during cancer development , 2003, British Journal of Cancer.

[38]  Z. Calay,et al.  Endometrial clear cell carcinoma metastatic to the paranasal sinuses: a case report and review of the literature. , 2004, Gynecologic oncology.

[39]  Jaw‐Ji Yang.  Mixed lineage kinase ZAK utilizing MKK7 and not MKK4 to activate the c-Jun N-terminal kinase and playing a role in the cell arrest. , 2002, Biochemical and biophysical research communications.

[40]  Amir Jazaeri,et al.  Microarray analysis reveals distinct gene expression profiles among different histologic types of endometrial cancer. , 2003, Cancer research.

[41]  J Wade Harper,et al.  The anaphase-promoting complex: it's not just for mitosis any more. , 2002, Genes & development.

[42]  C. Kallenberg,et al.  Molecular genetic analyses of human NKG2C (KLRC2) gene deletion. , 2004, International immunology.

[43]  M. Pepper.  Role of the Matrix Metalloproteinase and Plasminogen Activator-Plasmin Systems in Angiogenesis , 2001, Arteriosclerosis, thrombosis, and vascular biology.

[44]  V. Kosma,et al.  Expression of Matrix Metalloproteinase (MMP)-2 and MMP-9 in Breast Cancer with a Special Reference to Activator Protein-2, HER2, and Prognosis , 2004, Clinical Cancer Research.

[45]  Linda H. Shapiro,et al.  c-Maf Interacts with c-Myb To Regulate Transcription of an Early Myeloid Gene during Differentiation , 1998, Molecular and Cellular Biology.

[46]  Tsuyoshi Saito,et al.  Acceleration of invasive activity via matrix metalloproteinases by transfection of the estrogen receptor‐α gene in endometrial carcinoma cells , 2002, International journal of cancer.

[47]  Adam B. Olshen,et al.  Stratification of Intermediate-Risk Endometrial Cancer Patients into Groups at High Risk or Low Risk for Recurrence Based on Tumor Gene Expression Profiles , 2005, Clinical Cancer Research.

[48]  A. Berchuck,et al.  Microarray Analysis of Endometrial Carcinomas and Mixed Mullerian Tumors Reveals Distinct Gene Expression Profiles Associated with Different Histologic Types of Uterine Cancer , 2005, Clinical Cancer Research.

[49]  R. Barouki,et al.  The functional interaction between HMGA1 and the estrogen receptor requires either the N‐ or the C‐terminal domain of the receptor , 2004, FEBS letters.

[50]  A. El-Sohemy,et al.  Mevalonate Promotes the Growth of Tumors Derived from Human Cancer Cells in Vivo and Stimulates Proliferation in Vitro with Enhanced Cyclin-dependent Kinase-2 Activity* , 2004, Journal of Biological Chemistry.

[51]  R. Bentley,et al.  Malignant transformation of the human endometrium is associated with overexpression of lactoferrin messenger RNA and protein. , 1995, Cancer research.

[52]  L. Boxer,et al.  Repression of the c-myb Gene by WT1 Protein in T and B Cell Lines (*) , 1995, The Journal of Biological Chemistry.

[53]  E. Thompson,et al.  Doxycycline-Inducible Expression of SPARC/ Osteonectin/ BM40 in MDA-MB-231 Human Breast Cancer Cells Results in Growth Inhibition , 2002, Breast Cancer Research and Treatment.

[54]  Patrick Neven,et al.  Endometrial cancer , 2005, The Lancet.

[55]  K A Schafer,et al.  The Cell Cycle: A Review , 1998, Veterinary pathology.

[56]  K. Morgan,et al.  Gene regulation of the serine proteinase inhibitors α1-antitrypsin and α1-antichymotrypsin , 2001 .

[57]  J. Pollard,et al.  Progesterone Inhibits Estrogen-Induced Cyclin D1 and cdk4 Nuclear Translocation, Cyclin E- and Cyclin A-cdk2 Kinase Activation, and Cell Proliferation in Uterine Epithelial Cells in Mice , 1999, Molecular and Cellular Biology.

[58]  Qian Tao,et al.  The Stress-Responsive Gene GADD45G Is a Functional Tumor Suppressor, with Its Response to Environmental Stresses Frequently Disrupted Epigenetically in Multiple Tumors , 2005, Clinical Cancer Research.

[59]  David P. Kreil,et al.  Independent component analysis of microarray data in the study of endometrial cancer , 2004, Oncogene.

[60]  K. Morgan,et al.  Gene regulation of the serine proteinase inhibitors alpha1-antitrypsin and alpha1-antichymotrypsin. , 2002, Biochemical Society transactions.

[61]  R. Slebos,et al.  The frequency of p53, k‐ras mutations, and microsatellite instability differs in uterine endometrioid and serous carcinoma , 2000, Cancer.

[62]  H. Chung,et al.  Ursolic acid-induced down-regulation of MMP-9 gene is mediated through the nuclear translocation of glucocorticoid receptor in HT1080 human fibrosarcoma cells , 1998, Oncogene.

[63]  D. Kaufman,et al.  Lactoferrin: a tamoxifen-responsive protein in normal and malignant human endometrial cells in culture. , 2001, Experimental and molecular pathology.

[64]  Hui Zhang,et al.  Regulation of apoptosis at cell division by p34cdc2 phosphorylation of survivin. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[65]  M. Simons,et al.  Myb-dependent Regulation of Thrombospondin 2 Expression , 1998, The Journal of Biological Chemistry.

[66]  T. Löning,et al.  Expression of cell-cycle regulatory proteins in endometrial carcinomas: correlations with hormone receptor status and clinicopathologic parameters , 2001, Journal of Cancer Research and Clinical Oncology.