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Journal of Experimental Hematology ; (6): 1283-1288, 2017.
Article in Chinese | WPRIM | ID: wpr-301735


<p><b>OBJECTIVE</b>To investigate the effect and mechanism of miR-486 on glycometabolism of hematopoietic cells.</p><p><b>METHODS</b>qRT-PCR was applied to detect the expression of miR-486 or Sirt1 on TF-1 cells under hypoxia. Lentivirus was used to mediate the overexpression or inhibition of miR-486 on TF-1 cells and qRT-PCR was used to detect the expressions of Sirt1, glucose transporter 1(Glut1) and glucose transporter 4(Glut4). Lentivirus-mediated Sirt1-shRNA transduction was used to knockdown Sirt1 expression which was detected by qRT-PCR and Western blot. The expressions of Glut1 and Glut4 were determined by qRT-PCR.</p><p><b>RESULTS</b>Hypoxia promoted the expression of miR-486 and inhibited the expression of Sirt1. MiR-486 overexpression could inhibit the expression of Sirt1 and promote the expressions of Glut1 and Glut4, whereas miR-486 silencing upregulated the sirt1 expression and inhibited the expressions of Glut1 and Glut4. And inhibition of Sirt1 expression increased the expressions of Glut1 and Glut4.</p><p><b>CONCLUSION</b>MiR-486 can regulate the glycometabolism of hematopoietic cells by targeting Sirt1.</p>

Journal of Experimental Hematology ; (6): 1307-1311, 2009.
Article in Chinese | WPRIM | ID: wpr-343297


Mesenchymal stem cell (MSC)-based cell therapy has shifted into clinical trials to repair the damage of various tissues. In this setting, the survival of the transplanted cells contributes critically to the therapeutic effectiveness. To investigate the in vivo tracing of MSCs, a recombinant retroviral vector carrying firefly-luciferase reporter gene [pL (FLUC) SN] was constructed and several GPE+86 cell clones that stably expressed fluc were selected. The retroviral supernatants were collected and used to transfect MSC derived from C57 mice. The cells were then screened with G418 and the expression of the exogenous gene was identified by luciferase enzyme activity analysis. Labeled mouse MSCs (2x10(6)) were injected into skeletal muscles, and the in situ expression was noninvasively tracked by in vivo bioluminescence imaging for 1, 3 and 6 days after transplantation. The results showed that the survival rates of the grafted cells dropped sharply with time, they were 57.2+/-11.7%, 8.6+/-2.5% and 5.4+/-3.1% on day 1, 3 and 6 after transplantation, and no fluorescent signals above background were detected on day 10. It is concluded that the method described above could be used for in vivo tracing of grafted cells. Furthermore, MSCs could not survive even transplanted into the none-ischemic skeletal muscles.

Animals , Bone Marrow Cells , Cell Biology , Bone Marrow Transplantation , Methods , Cell Survival , Female , Genetic Vectors , Green Fluorescent Proteins , Luminescent Measurements , Methods , Mesenchymal Stem Cell Transplantation , Methods , Mesenchymal Stem Cells , Cell Biology , Mice , Mice, Inbred C57BL