Quantitative Proteomics Analysis of Altered Protein Expression in the Placental Villous Tissue of Early Pregnancy Loss Using Isobaric Tandem Mass Tags

Abstract

Many pregnant women suffer miscarriages during early gestation, but the description of these early pregnancy losses (EPL) can be somewhat confusing because of the complexities of early development. Thus, the identification of proteins with different expression profiles related to early pregnancy loss is essential for understanding the comprehensive pathophysiological mechanism. In this study, we report a gel-free tandem mass tags- (TMT-) labeling based proteomic analysis of five placental villous tissues from patients with early pregnancy loss and five from normal pregnant women. The application of this method resulted in the identification of 3423 proteins and 19647 peptides among the patient group and the matched normal control group. Qualitative and quantitative proteomic analysis revealed 51 proteins to be differentially abundant between the two groups (≥1.2-fold, Student's t-test, P < 0.05). To obtain an overview of the biological functions of the proteins whose expression levels altered significantly in EPL group, gene ontology analysis was performed. We also investigated the twelve proteins with a difference over 1.5-fold using pathways analysis. Our results demonstrate that the gel-free TMT-based proteomic approach allows the quantification of differences in protein expression levels, which is useful for obtaining molecular insights into early pregnancy loss.

Proteomics study reveals that the dysregulation of focal adhesion and ribosome contribute to early pregnancy loss

“…and Ni et al. , although the functions of most proteins are in common between our study and each of the other. The differences might be because we used TMT labeling based proteomics analysis, while Liu et al.…”

“…They found 51 differentially expressed proteins, of which 22 proteins were upregulated and 29 proteins were downregulated, including glutathione S‐transferase mu 2 (GSTM2), BC1 (ubiquinol‐cytochrome c reductase) synthesis‐like (BCS1L), and cullin 7 (CUL7). Moreover, they used pathway analysis and demonstrated that these differentially expressed proteins participated in cell migration, angiogenesis, oxidative stress, apoptosis, and metabolism pathways . Given the inconsistences between the abovementioned two studies, the molecular signaling pathways associated with EPL require further investigation.…”

Long-term effects of repeated superovulation on ovarian structure and function in rhesus monkeys

“…The aliquots were then combined, and the pooled samples were evaporated in a vacuum. Strong cation-exchange chromatography (SCX) fractionation was performed using an UltiMate 3000 high-pressure liquid chromatography (HPLC) system as previously described elsewhere (15)(16)(17). A total of 21 fractions were collected at 2-minute intervals over the SCX gradient and sequentially detected on an LTQ Orbitrap Velos (Thermo Fisher Scientific) using the data-dependent acquisition mode.…”

“…A total of 21 fractions were collected at 2-minute intervals over the SCX gradient and sequentially detected on an LTQ Orbitrap Velos (Thermo Fisher Scientific) using the data-dependent acquisition mode. The MS3 method was programmed as described in our previous studies (15)(16)(17).…”

The Use of Proteomics in Assisted Reproduction

“…In a more recent study, a gel-free TMT-labeling processing followed by LC-MS/MS proteomic of five placental villous tissues from women with early pregnancy loss has resulted in the identification of 51 differentially expressed proteins compared to normal placentas. The identified proteins were mainly involved in cell migration, RNA metabolism, cell differentiation, proliferation, metabolism and apoptosis, as well as reactive oxygen species generation and oxidative stress (129). Several proteomic studies have been conducted in search of early, fetal markers of aneuploidies.…”