title: In vitro culture significantly alters gene expression profiles and reduces differences between myometrial and fibroid smooth muscle cells.
abstract: Cultured myometrial (M) and fibroid (F) smooth muscle cells (SMCs) have been widely used as a model for the study of F growth. The aim of this study was to compare gene expression profiles using microarrays between six paired M and F tissues from hysterectomy specimens, as well as cells isolated from the same tissues and cultured for up to three passages. A total of 2055 genes were differentially expressed by ANOVA between all experimental groups. Among them, 128 genes were found to be statistically different between M and F tissues. More than 1100 genes were significantly changed between tissues and cultured cells, with 648 genes common between both M and F cells at P0 and P3. Expression profiles of six genes including estrogen receptor-alpha (ERalpha) and progesterone receptor (PR) were also validated using real-time PCR. These data demonstrate that large changes occur in SMC gene expression in culture, reducing differences between M and F cells. They also show that ERalpha and PR levels are reduced in cells compared with whole tissue. These results indicate that although M and F cell cultures provide an important tool to study these tumours, in vitro studies must be carefully planned and evaluated to provide meaningful results.
authors.affiliation: Centre for Women's Health Research, Monash University Department of Obstetrics and Gynaecology, Monash Institute of Medical Research, Clayton, Victoria, Australia.
authors.forename: Marina
authors.initials: M
authors.lastname: Zaitseva
authors.forename: Beverley J
authors.initials: BJ
authors.lastname: Vollenhoven
authors.forename: Peter A W
authors.initials: PA
authors.lastname: Rogers
mesh_terms: D002478:Cells, Cultured; D047628:Estrogen Receptor alpha / Q000235:genetics*; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D007889:Leiomyoma / Q000235:genetics / Q000473:pathology; D008875:Middle Aged; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D009215:Myometrium / Q000166:cytology / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D011980:Receptors, Progesterone / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
chemical_list: D047628:Estrogen Receptor alpha; D011980:Receptors, Progesterone