Human endometrium mRNA profile assessed by third dimensional oligonucleotide microarray analysis

Abstract

ObjectiveIn order to further comprehend the genetic events underlying the endometrial steroids response we undertook a 10.000 genes microarray analysis between human normal endometrium in the proliferative and secretory phases of the menstrual cycle.DesignThird dimensional microarray analysis.Materials and methodsHuman normal endometrial samples were obtained at the Gynecology Department, Pérola Byigton Hospital in São Paulo, Brazil. The patients were 80 women who were diagnosed as having leiomyomata in their uterus. Neither patient had received any hormonal treatment during the six months before the diagnosis of leiomyomata, endometrial fragments were divided in proliferative and secretory after histological analysis. Total RNA was extracted from samples by using TRIZOL (Invitrogen) according to manufacture`s instructions and further purified with Qiagen collums. RNA integrity was controlled on denaturing formaldehyde/agarose gel electrophoresis. Prior to reverse transcription, two pools of RNA were obtained by mixing 5 μg of total RNA from each individual case in two separated tubes representing endometrial tissues in proliferative (group 1) and secretory phase (group 2). These two pools were then hybridized two times each to four CodeLink UniSet Human I Bioarrays (Amersham Biosciences, Piscataway, NJ).Results257 genes were found differentially expressed between secretory and proliferative phases when a 5× difference was used as a cut-off criteria. Among them, genes involved in immune response, cell cycle control and calcium metabolism were the most strikingly differentially expressed.ConclusionGenes involved in immune response and calcium metabolism were found differentially expressed specially in the secretory phase. Authors hypothesize that overexpression of calcium related and immune responsive genes might be one of the hallmarks of the second phase of menstrual cycle. ObjectiveIn order to further comprehend the genetic events underlying the endometrial steroids response we undertook a 10.000 genes microarray analysis between human normal endometrium in the proliferative and secretory phases of the menstrual cycle. In order to further comprehend the genetic events underlying the endometrial steroids response we undertook a 10.000 genes microarray analysis between human normal endometrium in the proliferative and secretory phases of the menstrual cycle. DesignThird dimensional microarray analysis. Third dimensional microarray analysis. Materials and methodsHuman normal endometrial samples were obtained at the Gynecology Department, Pérola Byigton Hospital in São Paulo, Brazil. The patients were 80 women who were diagnosed as having leiomyomata in their uterus. Neither patient had received any hormonal treatment during the six months before the diagnosis of leiomyomata, endometrial fragments were divided in proliferative and secretory after histological analysis. Total RNA was extracted from samples by using TRIZOL (Invitrogen) according to manufacture`s instructions and further purified with Qiagen collums. RNA integrity was controlled on denaturing formaldehyde/agarose gel electrophoresis. Prior to reverse transcription, two pools of RNA were obtained by mixing 5 μg of total RNA from each individual case in two separated tubes representing endometrial tissues in proliferative (group 1) and secretory phase (group 2). These two pools were then hybridized two times each to four CodeLink UniSet Human I Bioarrays (Amersham Biosciences, Piscataway, NJ). Human normal endometrial samples were obtained at the Gynecology Department, Pérola Byigton Hospital in São Paulo, Brazil. The patients were 80 women who were diagnosed as having leiomyomata in their uterus. Neither patient had received any hormonal treatment during the six months before the diagnosis of leiomyomata, endometrial fragments were divided in proliferative and secretory after histological analysis. Total RNA was extracted from samples by using TRIZOL (Invitrogen) according to manufacture`s instructions and further purified with Qiagen collums. RNA integrity was controlled on denaturing formaldehyde/agarose gel electrophoresis. Prior to reverse transcription, two pools of RNA were obtained by mixing 5 μg of total RNA from each individual case in two separated tubes representing endometrial tissues in proliferative (group 1) and secretory phase (group 2). These two pools were then hybridized two times each to four CodeLink UniSet Human I Bioarrays (Amersham Biosciences, Piscataway, NJ). Results257 genes were found differentially expressed between secretory and proliferative phases when a 5× difference was used as a cut-off criteria. Among them, genes involved in immune response, cell cycle control and calcium metabolism were the most strikingly differentially expressed. 257 genes were found differentially expressed between secretory and proliferative phases when a 5× difference was used as a cut-off criteria. Among them, genes involved in immune response, cell cycle control and calcium metabolism were the most strikingly differentially expressed. ConclusionGenes involved in immune response and calcium metabolism were found differentially expressed specially in the secretory phase. Authors hypothesize that overexpression of calcium related and immune responsive genes might be one of the hallmarks of the second phase of menstrual cycle. Genes involved in immune response and calcium metabolism were found differentially expressed specially in the secretory phase. Authors hypothesize that overexpression of calcium related and immune responsive genes might be one of the hallmarks of the second phase of menstrual cycle.