Mol. Hum. Reprod. Advance Access originally published online on March 8, 2006
Molecular Human Reproduction 2006 12(3):187-207; doi:10.1093/molehr/gal018
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In vitro culture significantly alters gene expression profiles and reduces differences between myometrial and fibroid smooth muscle cells
Centre for Women’s Health Research, Monash University Department of Obstetrics and Gynaecology, Monash Institute of Medical Research, Clayton, Victoria, Australia
1 To whom correspondence should be addressed at: Centre for Women’s Health Research, Monash University Department of Obstetrics and Gynaecology, Monash Institute of Medical Research, 246 Clayton Road, Clayton, Victoria 3168, Australia. E-mail: marina.zaitseva{at}med.monash.edu.au
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-
(ER) 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 ER 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.
Key words:
fibroids/gene expression/in vitro culture/ER/PR
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