Microarray Analysis Of Differentially Expressed Genes In Cardinal Ligament From Women With Pelvic Organ Prolapse

●OBJECTIVES1.To identify the differentially expressed genes in cardinal ligament between patients with pelvic organ prolapse and those without POP by Human Genome Expression Chip.2.To further explore the possible molecular mechanism associated with pelvic organ prolapse(POP) based on the differential expression profile.●METHODS1.Cardinal ligament samples were obtained from three postmenopausal patients with POP-Q stageⅢand three age/BMI/parity matched postmenopausal subjects without POP.2.HE and Masson's trichrome staining was used to verify tissue origin and evaluate histological characteristics.3.Total RNA from samples were used to synthesize biotin-labeled cRNA by One-Cycle cDNA Synthesis method and hybridized with Human Genome Chip.4.The differentially expressed genes were identified between POP and normal controls with the threshold:Fold Change(FC)＞2.0 or＜0.5.The data was further interrogated with Gene Ontology(GO) and Pathway Analysis.5.Five upregulated genes were confirmed by quantitative reverse transcription-polymerase chain reaction.●RESULTS1.Alterations in ligament architecture seen in our population included disarrangement and collapse of smooth muscle bundles and collagen fibers. 2.The quality of total RNA was ensured by quantification and electrophoresis.The hybridization module of Human Genome expression chip was successfully established.3.179 differentially expressed genes were screened between POP and normal cardinal ligament tissue,including 20 function-unknown genes.107 genes were upregulated in POP group,while 72 downregulated.This differential expression profile was related to multiple functional proteins and pathways by biological analysis.Among these,Wnt signaling pathway was the most predominant.4.Quantitative PCR validated the upregulated expression of five genes.●CONCLUSIONS1.Ligament architecture in POP undergoes great alteration with disarrangement and collapse of smooth muscle bundles and collagen fibers.The mRNA elevations of COL1A1,COL3A1,COL5A1,MMP2 indicate an increase in collagen synthesis and breakdown.Degradation is predominant in collagen turnover and lead to ligament tissue defect.2.Genome Expression microarray is an effective approach for the high-throughput analysis of gene expression profile and aids to further explore the underlying molecular mechanism in POP.179 differentially expressed genes were screened between POP and normal cardinal ligament tissue,including 20 function-unknown genes.107 genes were upregulated in POP group,while 72 downregulated.3.The pathophysiology of POP is complex covering multiple functional proteins and pathways.Among these,Wnt signaling pathway was the most predominant and its antagonist DKK1,SFRP1 may contribute a neurodegenerative role in POP development.4.Quantitative PCR validated the upregulated expression of five genes from microarray data.