Exosome-mediated Hic-5 regulates proliferation and apoptosis of osteosarcoma via Wnt/β-catenin signal pathway
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Exosome-mediated Hic-5 regulates proliferation and apoptosis of osteosarcoma via Wnt/β-catenin signal pathway
HIC-5 expression was detected in osteosarkoma patients and osteosaroma cell lines by RT-PCR. Then RFP-SH-HIC-5 is transfected into the osteosarcoma cell line. The HIC-5 effect on cell viability, proliferation and apoptosis is assessed by MTT, Edu Kit and Flow Cytometry. Exosom is isolated from mg-63 cell supernatant by an exosomed insulation kit. Exosome-HIC-5 is confirmed by microscope electron transmission, particle size detection and RT-PCR.
Furthermore, the cells of the exosom-hic-5-5 are explored eligibility of cells, proliferation, and apoptosis. Furthermore, the C-IP test is used to identify the relationship between HIC-5 and SMAD4. TCF / LEF and the level of protein components of the WNT / β-Catenin signal are detected by the top luciferase test and Western blot. HIC-5 is regulated in osteosaroma tissue and cells. The decreased expression forced HIC-5 inhibits the proliferation of osteosarkoma cell lines, and induces the apoptosis of MG-63 and HOS. In Vivo, silencing HIC-5 sent the development of tumors. Furthermore, we isolate the exosom of the MG-63 supernatant, the excosom to conclude HIC-5 will regulate the proliferation and level of cell apoptosis of MG-63 and HOS. Furthermore, HIC-5 interacts with SMAD4 and regulates WNT / β-Catenin signals by reducing TCF / LEF activity.
Silencing HIC-5 inhibits proliferation and induces the apoptosis of cell osteosarcoma through deactivating the WNT / β-Catenin signal with the exosome path. Hypoxia is a characteristic of several serious diseases such as cardiovascular disorders and metabolism and cancer. A decrease at the oxygen level of the network induces hypoxic response in cells that seek to adapt to changing conditions. Failure to adapt to prolonged or severe hypoxia can trigger cell death. While some types of cells, such as neurons, are very susceptible to hypoxia, cancer cells utilize the hypoxia environment to undergo tumor growth, angiogenesis and metastasis. Hypoxic-induced processes trigger communication between complex cells and now there is an indication that the extracellular vesicle (EVS) plays a fundamental role in this process. The latest developments in EV isolation and the characterization methodology have raised awareness of the importance of EV purity in functional studies and cargo.
Exosomal limitations in the treatment of inflammatory diseases
Background: Despite great interest and many studies, there are currently no integrated standards that describe sequential manipulation with cells to get exosom for clinical use. The use of exosom has become an interesting alternative to cell therapy, because of the flexible properties of these biological vesicles. Enables scientists to manipulate their composition to produce the desired exosom to bring certain drugs, RNA and protein. This study aims to analyze the scientific literature on changes in the functional characteristics of exosom, depending on the manipulation method, which has the potential to contribute to the development of negative effects in the treatment of inflammatory genesis diseases. The choice of insulation method affects the set of protein markers disclosed, nucleic acid and receptors on microparticles.
Various surface receptors present on the exosome membrane can be engineered for the target of the lesion. Exocomals of healthy patients help reduce inflammation, normalize communication between cells and have antifibrotic, antioxidant and cytoprotective effects. Exosom can change the micro environment, but the micro environment can also change the composition of the exosomes.
Conclusion: Ecosomes obtained from sick patients carry markers of the characteristics of the appropriate disease. Such ecosomes can have pro-inflammatory properties, pro-fibrotic, cytotoxic, and oncogenic, and disrupt cellular cooperation. Until now, questions about doses, reactions to recurring giving, and the dose regime has not been fully resolved.
Exosome-mediated Hic-5 regulates proliferation and apoptosis of osteosarcoma via Wnt/β-catenin signal pathway
Proteomic exploration based on mass spectrometry of small urinary extracellular vesicles in vasculitis associated with ANCA compared to total urine
The vasculitis associated with the ANCA (AAV) is a rare, but potentially serious self-immune disease, if not nowadays, with increased mortality and morbidity. Finding the first activity biomarkers and the prognosis is very important. The small extracellular vesicles (EVS) isolated from the urine can be considered as a non-invasive source of biomarkers. We evaluated several protocols for the isolation of urinary electric vehicles.
To eliminate contaminating non-vesicular proteins due to AAV associated proteinuria, we used the processing of proteinase K. We have studied the differences in proteomes of small patient events with AAV compared to healthy controls by the freedom of LC-MS / MS labels. In parallel, we performed a similar proteomic analysis of urine samples from identical patients. The results of the study showed significant differences and similarities in the EV proteome and urine, the latter being strongly affected by proteinuria. Using bioinformatic tools, we have explored the modified proteins differently and their related routes by focusing on AAV’s pathophysiology. Our results indicate a significant regulation of Golgi enzymes, such as MAN1A1, which may be involved in the activation of the T cell by N-glycans glycosylation and can play a key role in the pathogenesis and diagnosis of AAV.
Description: Lipid droplets are lipid rich organelles found in the adipose tissue of eukaryotes. They function to regulate the hydrolysis and storage of neutral lipids and serve as storage for cholesterol and acyl-glycerols. Lipid droplets have also been associated with inflammatory responses, obesity, atherosclerosis, and cancer.
Description: The CD19 Positive Cell Isolation Kit is designed to magnetically separate CD19-expressing-cells from a complex immune cell population. This kit is optimized for the isolation of CD19 positive cells from normal human peripheral blood mononuclear cells (PBMCs). Cells are incubated with the antibody:bead complex and placed on a magnet for quick and easy separation. When placed on the magnet, CD19-positive cells will be immobilized along the side of the tube while undesired CD19-negative cells will remain in suspension for easy removal.
Description: The CD19 Positive Cell Isolation Kit is designed to magnetically separate CD19-expressing-cells from a complex immune cell population. This kit is optimized for the isolation of CD19 positive cells from normal human peripheral blood mononuclear cells (PBMCs). Cells are incubated with the antibody:bead complex and placed on a magnet for quick and easy separation. When placed on the magnet, CD19-positive cells will be immobilized along the side of the tube while undesired CD19-negative cells will remain in suspension for easy removal.
Meaning: The current study explores for the first time the changes in proteomes of small extracellular vesicles and urine of patients with vasculitis associated with former kidney relative to healthy controls by LC-MS / MS label. The isolation of vesicles from proteinuric urine samples has been modified to minimize plasma protein contamination and reduce co-insulation of extramuminal proteins. Differentially modified proteins and related routes with a role in AAV pathophysiology have been described and discussed. The results could be useful for the search for potential biomarkers in renal vasculitis associated with ANCa.
