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Journal of Experimental Hematology ; (6): 1531-1537, 2018.
Article in Chinese | WPRIM | ID: wpr-689902


<p><b>OBJECTIVE</b>To investigate the effects of exosomes derived from miR-486 gene-modified umbilical cord mesenchymal stem cells (UC-MSCs) on biological characteristics of rat cardiomyocytes.</p><p><b>METHODS</b>The human umbilical cord mesenchymal stem cells (UC-MSCs) were isolated and cultured, then the immunophenotypes and ability of osteogenic and adipogenic differentiation of UC-MSC were identified. The structure of exosomes was observed by electron microscopy; the effect of exosomes on cell migration was detected by transwell cell migration test; the miR-486 high expression of UC-MSC was mediated by using recombinant adenovirus vector, moreover the UC-MSC with high expression of miR-486 were identified by qPT-PCR. The exosomes were isolated from cell culture supernatant by ultracentrifugation and the miR-486 expression level of UC MSC exosomes was detected by qRT-PCR. The effect of exosomes on the proliferation of cardiomyocytes was evaluated by Dye670 marking. The HO-induced cardiomyocyte apoptosis model was established, and the effect of exosomes on apoptosis of cardiomyocytes was detected by flow cytometry with Annexin V/PI double staining.</p><p><b>RESULTS</b>The exosomes derived from UC-MSCs had the diameter between 40-100 nm and double membrane stracture. The recombinant adenovirus could effectively mediate the expression of miR-486 in UC-MSC, and the expression level of miR-486 in exosomes of miR-486-modified UC-MSC significantly increased. The exosomes with miR-486 high expression possessed the pro-proliferation and pro-migration effects on cardiomyocytes, moreover the preventive effect on apoptosis of cardiomyocytes.</p><p><b>CONCLUSION</b>The exosomes derived from UC-MSC and accompamied by high expression of miR-486 can promote the proliferation and migration of cardio myocytes, yet can prevent the apoptosis of cardiomyocytes.</p>

Journal of Experimental Hematology ; (6): 1267-1270, 2017.
Article in Chinese | WPRIM | ID: wpr-301738


Mesenchymal stem cells (MSC) possess important biological characteristics of tissue repair and regeneration. MSC exert the properties promoting endogenous angiogenesis and have been widely applied in treatment of ischemia diseases. The therapeutic potency of MSC for ischemia diseases is owing to their secretion of angiogenic growth factors and release of exosomes. MSC promote angiogenesis stronger in hypoxia environment, and their miRNA played an important role in mediating regulation. This review summarizes recent advances in treatment of angiogenesis using MSC and their mechanisms. The angiogenic activities of MSC under hypoxia condition and their regulation by a miRNA network were discussed.

Article in Chinese | WPRIM | ID: wpr-271885


<p><b>OBJECTIVE</b>To clarify the roles of SPK pathway in the regulation of proliferation, survival and glucose consume of human erythroleukemia TF-1 cells.</p><p><b>METHODS</b>The interfering in SPK expression of TF-1 cells was performed using leutivirus vector-mediated shRNA, the interference efficacy of SPK in TF-1 cells was detected by RT-qPCR and Western blot, the viability of TF-1 cell proliferation was detected by using CCK-8 method, the apoptosis of TF-1 cells was determined by flow cytmetry with Annexin V staining.</p><p><b>RESULTS</b>Hypoxia up-regulated the expression of HIF-1α, HIF-2α, and SPK in TF-1 cells. SPK treatment resulted in reduced proliferation and induced apoptosis in TF-1 cells. Furthermore, knockdown of the SPK significantly reduced utilization and consumption of glucose.</p><p><b>CONCLUSION</b>The SPK is key signalling molecule involved in regulation of hypoxia-induced proliferation and glucose metabolism in TF-1 cells, and plays an important rote in proliferation and energy metabolism of leukemia cells.</p>

Journal of Experimental Hematology ; (6): 1540-1544, 2014.
Article in Chinese | WPRIM | ID: wpr-340462


This study was aimed to explore the effect of VX-680, an aurora inhibitor, on proliferation and apoptosis of K562, KCL22 cell lines and CD34⁺ cells from chronic myeloid leukemia (CML) patients in vitro. The proliferation of K562 and KCL22 cell was detected by CCK-8 method. Apoptosis of cells was detected by Annexin V-PI labeling and flow cytometry. The colony forming ability of bone marrow CD34⁺ cells derived from CML patients and donors was determined by the colony forming test. The results showed that the treatment of K562, KCL22 and CML CD34⁺ cells with VX-680 of 20-100 nmol/L for 3 days could obviously inhibit the cell proliferation in a concentration-dependent manner (P < 0.01). VX-680 treatment significantly induced apoptosis of K562 and KCL22 cells. Compared to bone marrow CD34⁺ cells derived from the healthy donors, the colony forming ability of CML CD34⁺ cells derived from bone marrow of CML patients was remarkably reduced (P < 0.01). It is concluded that VX-680, an aurora inhibitor, can inhibit the proliferation and induce apoptosis of CML cells in vitro.

Apoptosis , Aurora Kinase A , Cell Line, Tumor , Cell Proliferation , Humans , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Pathology , Piperazines , Pharmacology , Protein Kinase Inhibitors , Pharmacology
Article in Chinese | WPRIM | ID: wpr-264957


The objective of this study was to explore the effects of microRNA-17-92 on the biological characteristics of K562 cells. The expression of miR-17-92 in K562 cells transfected with miRNA-17-92 mimic was detected by real time PCR. The effect of microRNA-17-92 on K562 cell proliferation was detected by CCK-8 method. Apoptosis of K562 cells was detected by Annexin V-PI labeling. Cell cycle distribution was determined by using flow cytometry. Western blot was performed to determine the protein levels of Crk. The results indicated that the transfection with miR-17-92 mimic increased expression of mature miR-17-92 in K562 cells. Compared with control group, cell proliferation and cell amount in S-phase of miR-17-92 mimic transfected group significantly increased, cell apoptosis decreased. The expression of signal connector protein Crk was greatly up-regulated in miR-17-92-mimic-transfected K562 cells. It is concluded that miR-17-92 can promote proliferation, inhibit apoptosis and regulate the cell cycle of K562 cells.

Apoptosis , Cell Cycle , Cell Proliferation , Gene Expression Regulation, Leukemic , HL-60 Cells , Humans , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Genetics , Metabolism , MicroRNAs , Genetics , Transfection
Article in Chinese | WPRIM | ID: wpr-686802


Case discussion is an effective approach,which combines basic theory with clinical medicine.It can evoke students' interest and cultivate their creative thinking capacity.Moreover,it can improve teachers' general ability in teaching pathophysiolo- gy.In this article,we discuss the application of case discussion in pathophysiology teaching.