Novel method to characterize primary cultures of leiomyoma and myometrium with the use of confirmatory biomarker gene arrays.
OBJECTIVE
To develop a rapid biomarker method for confirming that leiomyoma and myometrium primary cultures maintain the molecular phenotype of the progenitor tissues.


METHODS
Confirmation of primary cultures from leiomyoma and myometrium tissues.


METHODS
University hospital.


METHODS
Women undergoing hysterectomy for symptomatic leiomyomas.


METHODS
Primary cell cultures, reverse-transcriptase polymerase chain reaction (RT-PCR), microarray, real time RT-PCR, and immunofluorescence.


METHODS
Relative messenger RNA and protein expression in leiomyoma and myometrial cell cultures.


RESULTS
We developed primary cell cultures from human leiomyoma and patient-matched myometrium obtained from hysterectomy specimens. In the primary cultures, we confirmed the presence of smooth muscle-specific alpha-actin as well as filamentous actin. Based on microarray analysis, we expected and confirmed, in the progenitor tissue and derived primary cultures, an overexpression of versican (8.31 fold +/- 2.2 SEM and 4.3 fold +/- 1.01 SEM, respectively), transforming growth factor beta-3 (5.66 fold +/- 0.82 SEM and 4.92 fold +/- 0.58 SEM, respectively), and cytochrome P450-26A1 (6.76 fold +/- 0.80 SEM and 6.17 fold +/- 2.02 SEM, respectively), and an underexpression of dermatopontin (-5.6 fold +/- 1.82 SEM and -3.41 +/- 1.20 SEM, respectively).


CONCLUSIONS
Primary cell cultures offer a reliable in vitro model system for leiomyoma disease, if confirmed. Analysis of a gene array that distinguishes between myometrium and leiomyoma molecular phenotypes offers a rapid and reliable confirmation method, and provides confidence that in vitro findings resemble in vivo disease.
Malik
Minnie
M
Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814, USA.
Catherino
William H
WH
D018929:Cell Culture Techniques / Q000379:methods* / Q000639:trends; D002478:Cells, Cultured; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology; D005819:Genetic Markers; D006801:Humans; D007889:Leiomyoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D009215:Myometrium / Q000166:cytology* / Q000473:pathology* / Q000502:physiology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology
D005819:Genetic Markers