title: Fibroids display an anti-angiogenic gene expression profile when compared with adjacent myometrium.
abstract: The aetiology of uterine fibroids remains unknown, despite causing significant gynaecological morbidity. Fibroids have a reduced microvascular density when compared with adjacent myometrial tissue. The aim of this study was to identify genes with differential expression between fibroid and adjacent normal myometrium, particularly genes with a role in angiogenesis. Total RNA was extracted from fibroid/myometrium pairs from 12 hysterectomy specimens, and used to perform 24 cDNA microarrays. There were 10,500 genes screened on each microarray for differential expression. Analysis of expression data was carried out using multiple t-tests, as well as a novel class prediction algorithm (GeneRaVE). The differential gene expression of selected genes was confirmed by quantitative 'real time' RT-PCR. Selected genes with a role in angiogenesis were further analysed for expression in isolated cell populations of endothelial cells (fibroid and myometrium) and smooth muscle cells (fibroid and myometrium), to see if their expression was confined to particular cell types. Twenty-five genes with differential gene expression between fibroid and myometrium were identified. Insulin-like growth factor-2, endothelin A receptor, connective tissue growth factor (CTGF), cysteine-rich angiogenic inducer 61 (CYR61) and collagen 4alpha2 (COL4A2) were confirmed by RT-PCR. CTGF and CYR61, both angiogenesis promoters, were reduced in expression relative to myometrium. COL4A2, the precursor for an angiogenesis inhibitor, canstatin, was increased relative to myometrium. These three genes display an anti-angiogenic expression profile in fibroids relative to myometrium. These findings may explain the reduced microvascular density seen in fibroids relative to myometrium.
authors.affiliation: Centre for Women's Health Research, Monash University Department of Obstetrics & Gynaecology, 246 Clayton Rd, Melbourne, Australia. gareth.weston@med.monash.edu.au
authors.forename: Gareth
authors.initials: G
authors.lastname: Weston
authors.forename: Albert C
authors.initials: AC
authors.lastname: Trajstman
authors.forename: Caroline E
authors.initials: CE
authors.lastname: Gargett
authors.forename: Ursula
authors.initials: U
authors.lastname: Manuelpillai
authors.forename: Beverley J
authors.initials: BJ
authors.lastname: Vollenhoven
authors.forename: Peter A W
authors.initials: PA
authors.lastname: Rogers
mesh_terms: D000328:Adult; D020533:Angiogenesis Inhibitors / Q000235:genetics*; D001483:Base Sequence; D003094:Collagen / Q000235:genetics; D055513:Connective Tissue Growth Factor; D055515:Cysteine-Rich Protein 61; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics; D007889:Leiomyoma / Q000235:genetics*; D008875:Middle Aged; D009215:Myometrium / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D020533:Angiogenesis Inhibitors; C501848:CCN1 protein, human; C526317:CCN2 protein, human; D055515:Cysteine-Rich Protein 61; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D055513:Connective Tissue Growth Factor; D003094:Collagen