iTRAQ and PRM-based quantitative proteomics in early recurrent spontaneous abortion: biomarkers discovery

Cui, Ying; He, Ling; Yang, Chunyan; Ye, Qian

doi:10.1186/s12014-019-9256-y

Cited by 24 publications

(19 citation statements)

References 31 publications

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“…The PRM results were analyzed with Skyline quantitative analysis [43]. The expression of target proteins was obtained in the sample with the corrected internal standard peptide signal [44]. [46].…”

Section: Target Analysis By Parallel Reaction Monitoring (Prm)mentioning

confidence: 99%

Proteomic Analysis of Hypoxia-Induced Senescence of Human Bone Marrow Mesenchymal Stem Cells

Mai

Chen

et al. 2021

Stem Cells International

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Background and Aim. The senescence of human bone marrow mesenchymal stem cells (hBMSCs) can be induced by oxidative stress, but the mechanism by which it occurs is not yet clear. Here, we performed an investigation on the pathogenesis of hypoxia-induced senescence through proteomic analyses and aimed to explore the mechanisms of stem cell senescence. Methods. Hypoxia in hBMSCs was induced for 0, 4, and 12 hours, and cellular senescence was evaluated by senescence-associated β-galactosidase (SA-β-gal) staining. Tandem mass tag (TMT) labeling was combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) for differential proteomic analysis of hypoxia in hBMSCs. Parallel reaction monitoring (PRM) analysis was used to validate the candidate proteins. Verifications of signaling pathways were evaluated by western blotting. Cell apoptosis was evaluated using Annexin V/7-AAD staining by flow cytometry. The production of reactive oxygen species (ROS) was detected by the fluorescent probe 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA). Results. Cell senescence detected by SA-β-gal activity was higher in the 12-hour hypoxia-induced group. TMT analysis of 12-hour hypoxia-induced cells identified over 6000 proteins, including 686 differentially expressed proteins. Based on biological pathway analysis, we found that the senescence-associated proteins were predominantly enriched in the cancer pathways, PI3K-Akt pathway, and cellular senescence signaling pathways. CDK1, CDK2, and CCND1 were important nodes in PPI analyses. Moreover, the CCND1, UQCRH, and COX7C expressions were verified by PRM. Hypoxia induction for 12 hours in hBMSCs reduced CCND1 expression but promoted ROS production and cell apoptosis. Such effects were markedly reduced by the PI3K agonist, 740 Y-P, and attenuated by LY294002. Conclusions. Hypoxia of hBMSCs inhibited CCND1 expression but promoted ROS production and cell apoptosis through activating the PI3K-dependent signaling pathway. These findings provided a detailed characterization of the proteomic profiles related to hypoxia-induced senescence of hBMSCs and facilitated our understanding of the molecular mechanisms leading to stem cell senescence.

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Section: Target Analysis By Parallel Reaction Monitoring (Prm)mentioning

confidence: 99%

Proteomic Analysis of Hypoxia-Induced Senescence of Human Bone Marrow Mesenchymal Stem Cells

Mai

Chen

et al. 2021

Stem Cells International

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“…[19][20][21][22][23] Some signal amplification strategies based on RCA have been applied to achieve sensitive detection of markers at low levels. [24][25][26][27] Proteins are the key markers of disease, [28][29][30][31] therefore the development of highly sensitive methods to detect protein markers is a promising direction for disease. This paper reviews protein detection based on RCA using for example fluorescence [32] and electrochemistry.…”

Section: Introductionmentioning

confidence: 99%

Protein detection based on rolling circle amplification sensors

Shi

Cui

Sulemana³

et al. 2021

Luminescence

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Rolling circle amplification (RCA) is an isothermal process under the action of DNA polymerases. Large-scale DNA templates have been generated using RCA for target detection. Some signal amplification strategies including optical sensors and electrochemical sensors based on RCA have been applied to achieve sensitive detection.Sensors based on RCA have attracted increasing interest. Advances in RCA-based sensors for protein detection are reviewed in this paper. The advantages and detection mechanisms of sensors based on RCA are revealed and discussed. Finally, possible challenges and future perspectives are also outlined.

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“…Recurrent spontaneous abortion (RSA) is a common gynecological disease in China, de ned by two or more failed pregnancies, its etiology remains unclear, genetics, autoimmune abnormalities, endocrine, anatomy, or pre-thrombotic state are thought to be related to its pathogenesis [15,16]. Previous proteomic study had shown that there is a signi cant changes occurred in the pathway of Fc gamma R-mediated phagocytosis in RSA, this result indicated that autoimmunity is invloved in its pathogenesis [17]. A study suggested that RSA may be caused by an imbalance of vaginal ora (especially that of Pseudomonas) in RSA patients [18].…”

Section: Introductionmentioning

confidence: 99%

Gut Microbiota Composition and Functional Prediction in Recurrent Spontaneous Abortion

Cui

Zou

et al. 2021

Preprint

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Objective The changes of microbial community in pregnant women, let alone those of patients with recurrent spontaneous abortion (RSA), remain unclear. We analyzed the differences of gut mircobiota (GM) between RSA patients and pregnant women to find the possible mechanism of RSA. MethodsWe enrolled 30 RSA patients (RSA group) and 30 pregnant women who terminated their pregnancy and did not have a history of spontaneous abortion (NR group) in our hospital from June 2020 to August 2020, and fecal samples were obtained to analyze the GM using 16S rDNA V3–V4 sequencing.ResultsAt the phylum level, we found that there is no significant difference in composition of GM between RSA and NR. But at the genus level, compared with NR, Roseburia significantly decreased (P<0.01), and Ruminococcus significantly increased in RSA patients (P<0.05). Further analysis indicated that Klebsiella (P<0.05) was significantly increased, Prevotella.9 (P<0.05) and Roseburia (P<0.05) were significantly decreased in RSA2 group (BMI>23.9 in RSA). Moreover, Agathobacter (P<0.01) was significantly increased in NR2 group (no delivery in NR). Functional prediction indicated that GM may interfere with RSA through membrane transport, carbohydrate metabolism, amino acid metabolism and other pathways.ConclusionDecreased Roseburia in GM of pregnant women maybe related to RSA. Our results provide the basis for in-depth studies of the composition of gut microbial communities in RSA.

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iTRAQ and PRM-based quantitative proteomics in early recurrent spontaneous abortion: biomarkers discovery

Cited by 24 publications

References 31 publications

Proteomic Analysis of Hypoxia-Induced Senescence of Human Bone Marrow Mesenchymal Stem Cells

Proteomic Analysis of Hypoxia-Induced Senescence of Human Bone Marrow Mesenchymal Stem Cells

Protein detection based on rolling circle amplification sensors

Gut Microbiota Composition and Functional Prediction in Recurrent Spontaneous Abortion

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