Up regulation in gene expression of chromatin remodelling factors in cervical intraepithelial neoplasia

BackgroundThe highest rates of cervical cancer are found in developing countries. Frontline monitoring has reduced these rates in developed countries and present day screening programs primarily identify precancerous lesions termed cervical intraepithelial neoplasias (CIN). CIN lesions described as mild dysplasia (CIN I) are likely to spontaneously regress while CIN III lesions (severe dysplasia) are likely to progress if untreated. Thoughtful consideration of gene expression changes paralleling the progressive pre invasive neoplastic development will yield insight into the key casual events involved in cervical cancer development.ResultsIn this study, we have identified gene expression changes across 16 cervical cases (CIN I, CIN II, CIN III and normal cervical epithelium) using the unbiased long serial analysis of gene expression (L-SAGE) method. The 16 L-SAGE libraries were sequenced to the level of 2,481,387 tags, creating the largest SAGE data collection for cervical tissue worldwide. We have identified 222 genes differentially expressed between normal cervical tissue and CIN III. Many of these genes influence biological functions characteristic of cancer, such as cell death, cell growth/proliferation and cellular movement. Evaluation of these genes through network interactions identified multiple candidates that influence regulation of cellular transcription through chromatin remodelling (SMARCC1, NCOR1, MRFAP1 and MORF4L2). Further, these expression events are focused at the critical junction in disease development of moderate dysplasia (CIN II) indicating a role for chromatin remodelling as part of cervical cancer development.ConclusionWe have created a valuable publically available resource for the study of gene expression in precancerous cervical lesions. Our results indicate deregulation of the chromatin remodelling complex components and its influencing factors occur in the development of CIN lesions. The increase in SWI/SNF stabilizing molecule SMARCC1 and other novel genes has not been previously illustrated as events in the early stages of dysplasia development and thus not only provides novel candidate markers for screening but a biological function for targeting treatment.

[1]  G. Riggins,et al.  Characterization of the global profile of genes expressed in cervical epithelium by Serial Analysis of Gene Expression (SAGE) , 2005, BMC Genomics.

[2]  C. Cinti,et al.  Expression of cell-cycle-associated proteins pRB2/p130 and p27kip in vulvar squamous cell carcinomas. , 2001, Human pathology.

[3]  Alex E. Lash,et al.  Gene Expression Omnibus: NCBI gene expression and hybridization array data repository , 2002, Nucleic Acids Res..

[4]  Purvesh Khatri,et al.  Onto-Tools, the toolkit of the modern biologist: Onto-Express, Onto-Compare, Onto-Design and Onto-Translate , 2003, Nucleic Acids Res..

[5]  Martial Guillaud,et al.  Exploratory analysis of quantitative histopathology of cervical intraepithelial neoplasia: Objectivity, reproducibility, malignancy‐associated changes, and human papillomavirus , 2004, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[6]  K. Tominaga,et al.  MRGX Is a Novel Transcriptional Regulator That Exhibits Activation or Repression of the B-myb Promoter in a Cell Type-dependent Manner* , 2003, Journal of Biological Chemistry.

[7]  E. Knudsen,et al.  SWI/SNF Activity Is Required for the Repression of Deoxyribonucleotide Triphosphate Metabolic Enzymes via the Recruitment of mSin3B* , 2007, Journal of Biological Chemistry.

[8]  Y. Assaraf.  Molecular basis of antifolate resistance , 2007, Cancer and Metastasis Reviews.

[9]  J. Rader,et al.  Profiling microdissected epithelium and stroma to model genomic signatures for cervical carcinogenesis accommodating for covariates. , 2007, Cancer research.

[10]  Jianping Ding,et al.  The MRG domain of human MRG15 uses a shallow hydrophobic pocket to interact with the N‐terminal region of PAM14 , 2006, Protein science : a publication of the Protein Society.

[11]  C. Wheeler.  Advances in primary and secondary interventions for cervical cancer: human papillomavirus prophylactic vaccines and testing , 2007, Nature Clinical Practice Oncology.

[12]  A. Giordano,et al.  RB and cell cycle progression , 2006, Oncogene.

[13]  C. Woodman,et al.  The natural history of cervical HPV infection: unresolved issues , 2007, Nature Reviews Cancer.

[14]  Kenneth H Buetow,et al.  An anatomy of normal and malignant gene expression , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[15]  K. Basso,et al.  Gene dosage alterations revealed by cDNA microarray analysis in cervical cancer: Identification of candidate amplified and overexpressed genes , 2007, Genes, chromosomes & cancer.

[16]  Margaret A. Pericak-Vance,et al.  A comparative analysis of the information content in long and short SAGE libraries , 2006, BMC Bioinformatics.

[17]  Benyue Zhang,et al.  The E7 proteins of low- and high-risk human papillomaviruses share the ability to target the pRB family member p130 for degradation. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[18]  M. Matzuk,et al.  MRG15 Regulates Embryonic Development and Cell Proliferation , 2005, Molecular and Cellular Biology.

[19]  C. Cinti,et al.  Genetic alterations disrupting the nuclear localization of the retinoblastoma-related gene RB2/p130 in human tumor cell lines and primary tumors. , 2000, Cancer research.

[20]  P. Good,et al.  Permutation Tests: A Practical Guide to Resampling Methods for Testing Hypotheses , 1995 .

[21]  F. Quiocho,et al.  Multipurpose MRG domain involved in cell senescence and proliferation exhibits structural homology to a DNA-interacting domain. , 2006, Structure.

[22]  C. Simone,et al.  SWI/SNF: The crossroads where extracellular signaling pathways meet chromatin , 2006, Journal of cellular physiology.

[23]  W. Muller,et al.  The phosphatidyl inositol 3-kinase signaling network: implications for human breast cancer , 2007, Oncogene.

[24]  M. Matzuk,et al.  MrgX Is Not Essential for Cell Growth and Development in the Mouse , 2005, Molecular and Cellular Biology.

[25]  S. Franceschi,et al.  Human papillomavirus and HPV vaccines: a review. , 2007, Bulletin of the World Health Organization.

[26]  J. Torchia,et al.  A Novel Nuclear Receptor Corepressor Complex, N-CoR, Contains Components of the Mammalian SWI/SNF Complex and the Corepressor KAP-1* , 2000, The Journal of Biological Chemistry.

[27]  D. Ayer,et al.  Role for the Mortality Factors MORF4, MRGX, and MRG15 in Transcriptional Repression via Associations with Pf1, mSin3A, and Transducin-Like Enhancer of Split , 2002, Molecular and Cellular Biology.

[28]  C. Boerkoel,et al.  Advances in chromatin remodeling and human disease. , 2004, Current opinion in genetics & development.

[29]  Yusuke Nakamura,et al.  Inhibition of Experimental Intimal Thickening in Mice Lacking a Novel G-Protein–Coupled Receptor , 2003, Circulation.

[30]  E. Lees,et al.  Cyclin E Associates with BAF155 and BRG1, Components of the Mammalian SWI-SNF Complex, and Alters the Ability of BRG1 To Induce Growth Arrest , 1999, Molecular and Cellular Biology.

[31]  G. Tortolero-Luna,et al.  Human papillomavirus infection: biology, epidemiology, and prevention , 2005, International Journal of Gynecologic Cancer.

[32]  R. Seong,et al.  SRG3 Interacts Directly with the Major Components of the SWI/SNF Chromatin Remodeling Complex and Protects Them from Proteasomal Degradation* , 2007, Journal of Biological Chemistry.

[33]  J. Rowley,et al.  Identifying novel transcripts and novel genes in the human genome by using novel SAGE tags , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[34]  S. Namkoong,et al.  Absence of p15INK4B and p16INK4A gene alterations in primary cervical carcinoma tissues and cell lines with human papillomavirus infection. , 1998, Gynecologic Oncology.

[35]  J. Lee,et al.  Expression of PTEN in the progression of cervical neoplasia and its relation to tumor behavior and angiogenesis in invasive squamous cell carcinoma , 2006, Journal of surgical oncology.

[36]  C. Cinti,et al.  Mutations in the retinoblastoma-related gene RB2/p130 in primary nasopharyngeal carcinoma. , 2000, Cancer research.

[37]  B. Gusterson,et al.  E7 proteins from oncogenic human papillomavirus types transactivate p73: role in cervical intraepithelial neoplasia , 2002, British Journal of Cancer.

[38]  M. Matzuk,et al.  MRG 15 Regulates Embryonic Development and Cell Proliferation , 2005 .

[39]  S. Namkoong,et al.  Absence ofp15INK4Bandp16INK4AGene Alterations in Primary Cervical Carcinoma Tissues and Cell Lines with Human Papillomavirus Infection , 1998 .

[40]  O. Pereira-smith,et al.  MRG15 Activates the B-myb Promoter through Formation of a Nuclear Complex with the Retinoblastoma Protein and the Novel Protein PAM14* , 2001, The Journal of Biological Chemistry.

[41]  Ji Huang,et al.  [Serial analysis of gene expression]. , 2002, Yi chuan = Hereditas.

[42]  Calum MacAulay,et al.  Whole genome tiling path array CGH analysis of segmental copy number alterations in cervical cancer cell lines , 2007, International journal of cancer.

[43]  R. Watson,et al.  Cell cycle regulation by the B-Myb transcription factor , 2003, Cellular and Molecular Life Sciences CMLS.

[44]  C. Cinti,et al.  Expression of cell-cycle–associated proteins pRB2/p130 and p27kip1 in vulvar squamous cell carcinomas , 2001 .

[45]  R. Roberts,et al.  Human Genome and Diseases:¶WD-repeat proteins: structure characteristics, biological function, and their involvement in human diseases , 2001, Cellular and Molecular Life Sciences CMLS.

[46]  D. Hanahan,et al.  The Hallmarks of Cancer , 2000, Cell.

[47]  C. MacAulay,et al.  Comprehensive serial analysis of gene expression of the cervical transcriptome , 2007, BMC Genomics.

[48]  P. Khatri,et al.  Global functional profiling of gene expression ? ? This work was funded in part by a Sun Microsystem , 2003 .