Platelet miRNA expression from patients of PDAC and healthy donors
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ABSTRACT:
PROVIDER: PRJNA838609 | ENA |
REPOSITORIES: ENA
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Project description:To identify the miRNA expressing profiles of Platelet microparticles（PMPs, we have employed the Agilent Human miRNA 8×60K (Design ID:046064) microarray. Platelet microparticles. The platelets were derived from citrated blood of healthy human donors under an Institutional Review Board-approved protocol. Platelets were isolated after centrifugation of blood (1200r for 30 min at 21℃), then the supernatant (platelet-rich plasma) was centrifuged at 2000r for 30 min at 21℃, and the pellet containing platelets was resuspended in RPMI-1640 medium (HyClone, Logan, UT). Platelets were counted (Clinical Laboratory, Shanghai First Maternity and Infant Hospital, Shanghai) and adjusted to a density of 150 × 106 cells/mL before supplement with 1.5% ACD(sigma ) and stimulated with thrombin (1.0 u/mL; Takeda Austria) for 1 h. PMPs were in the supernatant after centrifugation at 4000r for 10 min at 4℃,then the supernatants were centrifuged at 50,000 × g for 60 min at 4 °C. The pellets containing MPs were resuspended in RPMI-1640 medium and quantified by BCA method.
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Project description:Upon activation, platelets release a multitude of soluble and vesicular signals, collectively termed the ‘platelet releasate’ (PR). This PR plays an important role in haemostasis, wound healing and the inflammatory response. We and others have used qualitative/quantitative proteomic approaches to characterise the PR; however, with reports of marked inter-individual variability, confident and reliable insights have been hindered. Here we aimed to provide a reproducible and quantifiable proteomic analysis and establish a 'core' human PR, as well as to assess its variability in healthy human pregnancy where increased platelet activation is observed. 896 proteins from thrombin-induced PRs were quantified across 32healthy donors, with 277 proteins forming a ‘core’ PR with low variability. All 277 core PR proteins were identified in pregnancy. Furthermore 69 proteins were found differentially released from platelets in healthy human pregnancy. In summary, the PR comprises a ‘core’ set of proteins between healthy individuals. This dynamic proteome significantly changes in physiologic and pathologic conditions. Thus, the PR is a barcode for the health status of an individual at a given time.
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