Global hypomethylation of genomic DNA in cancer-associated myofibroblasts.

Global hypomethylation has long been recognized as a feature of the malignant epithelial component in human carcinomas. Here we show evidence for this same type of epigenetic alteration in cancer-associated stromal myofibroblasts. We used methylation-sensitive SNP array analysis (MSNP) to profile DNA methylation in early-passage cultures of stromal myofibroblasts isolated from human gastric cancers. The MSNP data indicated widespread hypomethylation in these cells, with rare focal gains of methylation, conclusions that were independently validated by bisulfite sequencing and by a methylation-sensitive cytosine incorporation assay. Immunohistochemistry with anti-5-methylcytosine (anti-5-methyl-C) in a series of gastrectomy specimens showed frequent loss of methylation in nuclei of both the malignant epithelial cells and alpha-smooth muscle actin (ASMA)-positive stromal myofibroblasts of both intestinal-type and diffuse carcinomas. We confirmed this phenomenon and established its onset at the stage of noninvasive dysplastic lesions by immunohistochemistry for anti-5-methyl-C in a transgenic mouse model of multistage gastric carcinogenesis. These findings indicate similar general classes of epigenetic alterations in carcinoma cells and their accompanying reactive stromal cells and add to accumulating evidence for biological differences between normal and cancer-associated myofibroblasts.

[1]  B. Tycko,et al.  Genomic surveys by methylation-sensitive SNP analysis identify sequence-dependent allele-specific DNA methylation , 2008, Nature Genetics.

[2]  Seung‐Mo Hong,et al.  Pancreatic cancer associated fibroblasts display normal allelotypes , 2008, Cancer biology & therapy.

[3]  D. Roukos.  Breast-cancer stromal cells with TP53 mutations. , 2008, The New England journal of medicine.

[4]  P. Platzer,et al.  Breast-cancer stromal cells with TP53 mutations and nodal metastases. , 2007, The New England journal of medicine.

[5]  Kathleen R. Cho,et al.  Mouse model of colonic adenoma-carcinoma progression based on somatic Apc inactivation. , 2007, Cancer research.

[6]  D. V. Von Hoff,et al.  Tumor-stroma interactions in pancreatic ductal adenocarcinoma , 2007, Molecular Cancer Therapeutics.

[7]  M. Monden,et al.  Stromal Myofibroblasts Predict Disease Recurrence for Colorectal Cancer , 2007, Clinical Cancer Research.

[8]  A. Thorne,et al.  Regulation of myofibroblast transdifferentiation by DNA methylation and MeCP2: implications for wound healing and fibrogenesis , 2007, Cell Death and Differentiation.

[9]  Robert A. Weinberg,et al.  Stromal Fibroblasts in Cancer: A Novel Tumor-Promoting Cell Type , 2006, Cell cycle.

[10]  K. Bodger,et al.  The role of matrix metalloproteinase-7 in redefining the gastric microenvironment in response to Helicobacter pylori. , 2006, Gastroenterology.

[11]  B. Tycko,et al.  A single nucleotide polymorphism chip-based method for combined genetic and epigenetic profiling: validation in decitabine therapy and tumor/normal comparisons. , 2006, Cancer research.

[12]  Kentaro Yamashita,et al.  Global DNA demethylation in gastrointestinal cancer is age dependent and precedes genomic damage. , 2006, Cancer cell.

[13]  J. Gillespie,et al.  Gene promoter methylation in prostate tumor-associated stromal cells. , 2006, Journal of the National Cancer Institute.

[14]  B. Tycko,et al.  Genomic Profiling Maps Loss of Heterozygosity and Defines the Timing and Stage Dependence of Epigenetic and Genetic Events in Wilms' Tumors , 2005, Molecular Cancer Research.

[15]  Jun Yao,et al.  Distinct epigenetic changes in the stromal cells of breast cancers , 2005, Nature Genetics.

[16]  P. Adegboyega,et al.  Epithelial cells and their neighbors I. Role of intestinal myofibroblasts in development, repair, and cancer. , 2005, American journal of physiology. Gastrointestinal and liver physiology.

[17]  K. Andersson.  Alpha1 adrenoceptor subtypes and bladder function. , 2005, The Journal of urology.

[18]  C. Morrison,et al.  Combined Total Genome Loss of Heterozygosity Scan of Breast Cancer Stroma and Epithelium Reveals Multiplicity of Stromal Targets , 2004, Cancer Research.

[19]  Giulio Gabbiani,et al.  The stroma reaction myofibroblast: a key player in the control of tumor cell behavior. , 2004, The International journal of developmental biology.

[20]  A. Balmain,et al.  A Mouse Skin Multistage Carcinogenesis Model Reflects the Aberrant DNA Methylation Patterns of Human Tumors , 2004, Cancer Research.

[21]  Cheng Li,et al.  dChipSNP: significance curve and clustering of SNP-array-based loss-of-heterozygosity data , 2004, Bioinform..

[22]  A. Feinberg,et al.  The history of cancer epigenetics , 2004, Nature Reviews Cancer.

[23]  A. Kaneda,et al.  Frequent hypomethylation in multiple promoter CpG islands is associated with global hypomethylation, but not with frequent promoter hypermethylation , 2004, Cancer science.

[24]  R. Paterson,et al.  Molecular genetic alterations in the laser‐capture–microdissected stroma adjacent to bladder carcinoma , 2003, Cancer.

[25]  M. Washington,et al.  Helicobacter pylori strain-selective induction of matrix metalloproteinase-7 in vitro and within gastric mucosa. , 2003, Gastroenterology.

[26]  M. Goggins,et al.  Effects of 5-aza-2'-deoxycytidine on matrix metalloproteinase expression and pancreatic cancer cell invasiveness. , 2003, Journal of the National Cancer Institute.

[27]  Long-Cheng Li,et al.  MethPrimer: designing primers for methylation PCRs , 2002, Bioinform..

[28]  H. Nakayama,et al.  α Smooth muscle actin positive stromal cells in gastric carcinoma , 2002, Journal of clinical pathology.

[29]  M. Ehrlich,et al.  DNA methylation in cancer: too much, but also too little , 2002, Oncogene.

[30]  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.

[31]  D. Heimburger,et al.  Altered global methylation of DNA: an epigenetic difference in susceptibility for lung cancer is associated with its progression. , 2001, Human pathology.

[32]  C Eng,et al.  Excessive CpG island hypermethylation in cancer cell lines versus primary human malignancies. , 2001, Human molecular genetics.

[33]  A. Niveleau,et al.  Evaluation of global DNA hypomethylation in human colon cancer tissues by immunohistochemistry and image analysis , 2000, Gut.

[34]  G. Bratthauer,et al.  Concurrent and independent genetic alterations in the stromal and epithelial cells of mammary carcinoma: implications for tumorigenesis. , 2000, Cancer research.

[35]  D. Heimburger,et al.  Immunohistochemical Evaluation of Global DNA Methylation: Comparison with in Vitro Radiolabeled Methyl Incorporation Assay , 2000, Biotechnic & histochemistry : official publication of the Biological Stain Commission.

[36]  I. Pogribny,et al.  A sensitive new method for rapid detection of abnormal methylation patterns in global DNA and within CpG islands. , 1999, Biochemical and biophysical research communications.

[37]  J. Mason,et al.  Colonic mucosal concentrations of folate correlate well with blood measurements of folate status in persons with colorectal polyps. , 1998, The American journal of clinical nutrition.

[38]  L. Glória,et al.  Global DNA hypomethylation occurs in the early stages of intestinal type gastric carcinoma. , 1996, Gut.

[39]  W. Gerald,et al.  Epigenetic lesions at the H19 locus in Wilms' tumour patients , 1994, Nature Genetics.

[40]  H. Dvorak.  Tumors: wounds that do not heal. Similarities between tumor stroma generation and wound healing. , 1986, The New England journal of medicine.

[41]  M. Ehrlich,et al.  The 5-methylcytosine content of DNA from human tumors. , 1983, Nucleic acids research.

[42]  V. L. Wilson,et al.  DNA methylation decreases in aging but not in immortal cells. , 1983, Science.

[43]  A. Feinberg,et al.  Hypomethylation distinguishes genes of some human cancers from their normal counterparts , 1983, Nature.

[44]  Martin Widschwendter,et al.  Breast cancer DNA methylation profiles in cancer cells and tumor stroma: association with HER-2/neu status in primary breast cancer. , 2006, Cancer research.

[45]  L. Chung,et al.  Molecular insights into prostate cancer progression: the missing link of tumor microenvironment. , 2005, The Journal of urology.

[46]  L. Matrisian,et al.  Expression and localization of the matrix metalloproteinase pump‐1 (MMP‐7) in human gastric and colon carcinomas , 1991, Molecular carcinogenesis.