Abstract��
OBJECTIVE To explore the effects of sodium nitrite pretreatment on the chemotherapeutic sensitivity to adriamycin (ADM) in MG63 cells. METHODS MTT assay was used to measure the IC50 of ADM and the resistance index to MG-63 cells. FITC-annexin��/PI staining flow cytometry was used to analyze the apoptotic rate of cells. Cell death rate was assessed by PI and hochest33258 viable cell staining. The effects of autophagy in cells were investigated with EGFP-LC3b transfection and electron microscope. The expression of HIF-1�� and LC3 in cells was detected by Western blotting. RESULTS Compared to ADM treatment alone, MG-63 cells was pretreated with sodium nitrite(10-30 mg��L-1) for 24 h and followed exposed to ADM (0.31 ��g��mL-1) for 24 h,the IC50 of ADM and the resistance index to MG-63 increased significantly. These phenomenon could be reversed through down-regulation of autophagy with 3- methyladenine(3-MA). LC3-�� green dot fluorescence diffusely distributed near the cell membrane after treatment with ADM and the autophagosome could be found under the electronmicroscopy. ADM could induce autophagy and apoptosis, 3-MA could suppress autophagy, but had no significant effects on apoptosis. Compared to ADM treatment alone, MG-63 cells were pretreated with sodium nitrite (10 mg��L-1) for 24 h, and followed exposed to ADM, the autophagy-positive cells were increased significantly,3-MA administration following ADM could increase apoptosis and death rate in comparison to ADM treatment alone. In addition, western blotting showed that the expression of hypoxia-inducible factor 1��(HIF-1��) and LC3��/ LC3��in the cells that pretreated with sodium nitrite and followed exposed to ADM were higher than ADM treatment alone group. CONCLUSION Sodium nitrite pretreatment could decrease the chemotherapeutic sensitivity to ADM in osteosarcoma cells and the mechanism may be related to promote autophagy and increase the expression of HIF-1��.
MA Kun-,
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GUO Jian-Gang- etc
.Sodium Nitrite Pretreatment Decreases the Chemotherapeutic Sensitivity to Adriamycin in Osteosarcoma Cells[J] Chinese Pharmaceutical Journal, 2014,V49(16): 1397-1403
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