Mouse Anti-Human FN1 Hybridoma [Q4OQ/QGo]

Objective: Obstructive sleep apnea hypopnea syndrome (OSAHS) is an independent risk factor for many clinical complications. However, how OSAHS cause multiple organ injury and initiate inter-organ communication remains unclear. Moreover, despite it is well-recognized that chronic intermittent hypoxia (CIH) is a main feature of OSAHS, specific contribution of CIH to overall OSAHS-initiated pathological complications remains unclear. This study aimed to use an unbiased proteomic approach to determine whether OSAHS alters protein profiles of serum extracellular microvesicles (SEMVs) and how CIH contributes to such alterations. Methods: Tandem mass tag (TMT)-labeled quantitative proteomics assay was used to compare the differentially expressed proteins (DEPs) in SEMVs of OSAHS patients and non-OSAHS subjects. To evaluate the contribution of CIH to OSAHS, CIH rodent model was constructed and the same comparative proteomics study was performed in SEMVs from CIH and normoxia rats. The similarity and disparity of DEPs and DEPs-related functions predicted by bioinformatics tools were compared in above-mentioned two models, and several DEPs were selected and further verified by ELISA or Western blotting. Results: TMT-labeled quantitative proteomics assay unravels 32 DEPs in OSAHS patient SEMVs from a total of 560 human SEMV proteins identified. Four DEPs, namely C-reactive protein (CRP), Haptoglobin (HP),Fibronectin (FN1) and Platelet factor 4 (PF4), were further verified by ELISA and three of them (CRP, FN1 and Hp) showed significant difference in expression level between OSAHS and non-OSAHS groups. In SEMVs of rat CIH model, 121 DEPs out of 723 proteins were identified. By comparing the DEPs identified from the two models. 3 proteins (CRP and FN1 and F13a1) were found identical with the same alteration pattern (CRP was upregulated, FN1 and Fl3a1 were downregulated) in SEMVs from OSAHS patients and CIH rats, which were further verified by Western blotting. Computational functional analysis further revealed the common and distinct DEP-involved pathways under OSAHS or CIH status. Conclusions: This study provides the first evidence that OSAHS causes significant alteration in SEMV protein composition, which may contribute to OSAHS-triggered multiple organ injury and organ-to-organ communication. Moreover, we have demonstrated that CIH is the primary contributor for increased inflammatory protein expression in SEMV. As CRP is being increasingly recognized not only as a marker but also a mediator of inflammatory response to tissue injury, increased SEMV CRP in CIH/OSAHS may play an important role in OSAHSinduced tissue injury, suggesting SEMV CRP might be a therapeutic target against OSAHS-related complications. (C) 2018 Elsevier Inc. All rights reserved.

Considering the resistance of papillary thyroid cancer (PTC) I-131 therapy, this study was designed to find a solution at molecular respect. By probing into lncRNA-NEAT1/miR-101-3p/FN1 axis and PI3K/AKT signaling pathway, this study provided a potential target for PTC therapy. I-131-resistant cell lines were established by continuous treatment with median-lethal I-131. Bioinformatic analysis was applied to filtrate possible lncRNA/miRNA/mRNA and related signaling pathway. Luciferase reporter assay was employed in the verification of the targeting relationship between lncRNA and miRNA as well as miRNA and mRNA. MTT assay and flow cytometry assay were performed to observe the impact of NEAT1/miR-101-3p/FN1 on cell viability and apoptosis in radioactivity iodine (RAI)-resistant PTC cell lines, respectively. Western blot and qRT-PCR were conducted to measure the expression of proteins and mRNAs in RAI-resistant PTC tissues and cells. Meanwhile, endogenous PTC mice model were constructed, in order to verify the relation between NEAT1 and RAI-resistance in vivo. NEAT1 was over-expressed in RAI-resistant PTC tissues and cell lines and could resist RAI by accelerating proliferation accompanied by suppressing apoptosis. It indicated that overexpressed NEAT1 restrained the damage of RAI to tumor in both macroscopic and microcosmic. Besides, NEAT1/miR-101-3p exhibited a negative correlation by directly targeting each other. The expression of FN1, an overexpressed downstream protein in RAI-resistance PTC tissues, could be tuned down by miR-101-3p, while the decrease could be restored by NEAT1. In conclusion, both in vitro and in vivo, NEAT1 suppression could inhibit I-131 resistance of PTC by upregulating miR-101-3p/FN1 expression and inactivated PI3K/AKT signaling pathway both in vitro and in vivo.