Abstract
During labor, a variety of coordinated physiological and biochemical events cause the myometrium to transition from a quiescent to contractile state; the molecular mechanisms responsible for this transition, however, remain unclear. To better understand this transition at a molecular level, the global transcriptome and proteome of human myometrial samples in labor and those not in labor were investigated through RNA sequencing (RNA-seq) and quantitative liquid chromatography–tandem mass spectrometry (LC-MS/MS) via data-independent acquisition (DIA) and parallel reaction monitoring (PRM) methods. Furthermore, an integrated proteotranscriptomic analysis was performed to explore biological processes and pathway alterations during labor; this analysis identified 1,626 differentially expressed mRNAs (1,101 upregulated, 525 downregulated) and 135 differentially expressed proteins (97 upregulated, 38 downregulated) in myometrium between nonlabor and in labor groups. The comprehensive results of these analyses showed that the upregulated mRNAs and proteins increased inflammation under hypoxia stress in the myometrium under labor, and related proteins and cytokines were validated by PRM and Luminex assays. Our study confirmed the biological process of inflammation and hypoxia in laboring myometrium at the transcriptome and proteome levels and provided recourse to discover new molecular and biological changes during labor.
Highlights
IntroductionThe uterus, especially the myometrium, remains quiescence. during labor, the myometrium undergoes biochemical and structural changes, transforming from a relaxed, noncontractile phenotype to a highly coordinated and intensely contractile one
During human pregnancy, the uterus, especially the myometrium, remains quiescence
To establish a thorough transcriptome profile of NL and IL myometrium, we conducted an investigation through RNA sequencing (RNA-seq) on mRNA separated from the myometrium of 10 NL and 10 IL
Summary
The uterus, especially the myometrium, remains quiescence. during labor, the myometrium undergoes biochemical and structural changes, transforming from a relaxed, noncontractile phenotype to a highly coordinated and intensely contractile one. The transition of a quiescent myometrium state to an active contractive one requires complex and highly regulated changes in gene coding to drive changes in the structure, contractility, and signaling in the uterine myometrium [3, 4]. Stanfield et al integrated three transcriptome datasets in the NCBI Gene Expression Omnibus database to characterize 126 labor-associated genes; labor signatures include interleukins, cytokines, apoptosis, MYC, and cell proliferation/differentiation, whereas cyclic AMP signal and muscle relaxation pathway were mainly associated with nonlabor [6]. These signatures accurately categorize and describe the various stages of delivery. Despite the generation of some muscular transcriptome datasets, reliable human delivery transcriptional signals and pathway networks have yet to be studied; extensive and consistent studies and review comparisons are lacking
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