Gene Expression Analysis between Uterine Leiomyoma and Normal Myometrial Tissues by DNA Chip

Kwon, Sang Hoon; Cho, Chi Hum; Cha, Soon Do; Back, Won Ki; Kim, Moon Kyu; Kim, Jung Chul

Abstract

OBJECTIVE
To investigate the difference of gene expressions between leiomyoma and normal myometrial tissue was analzed by DNA Chip.
METHODS
cDNAs retro-transcribed from equal quantities of mRNA derived from leiomyoma and corresponding normal myometrial tissue were labeled with Cy5 and Cy3 fluorescein as probes. The mixed probe was hybridized with two pieces of 3,066 double dot from a human dermal papilla cell cDNA library and scanned with a laser scanner. The acquired image was analyzed by ImaGene 3.0 software. Validation of gene expression was performed by reverse transcription-Polymerase chain reaction (RT-PCR) in 5 leiomyomas and corresponding normal myometrial tissues.
RESULTS
Among many differentially expressed genes, genes with expression levels more than 3 times were found by comparing leiomyoma with corresponding normal myometrial tissue. One gene with expression levels lesser than 3 times in leiomyoma tissue compared to normal myometrial tissue was also detected. Although alterations of several genes, such as osteoblast specific factor 2, PAI-1 mRNA-binding protein, hydroxyacyl-Coenzyme A dehydrogenase/3-ketoacyl-Coenzyme A thiolase/enoyl-Coenzyme A hydratase alpha subunit (HADHA), p311, DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide 1 (DDX1), Hexokinase 1, 2 were identified in a significant high fraction of uterine leiomyoma compared to normal myometrial tissue. Cyr61 gene was shown to be markedly down-regulated in leiomyoma compared with the matched uterine myometrial control. I validated differential expression of genes by RT-PCR and demonstrated overexpression of OSF-2, HADHA, p311, DDX1, Hexokinase 1, 2.
CONCLUSION
DNA chip techniques are effective in screening differential gene expression between leiomyoma tissue and normal myometrial tissue. These genes may be related to the genesis and development of uterine leiomyoma. Analysis of the human leiomyoma gene expression profile by DNA chip may be helpful gene diagnosis, treatment and prevention of this disease.