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目的:观察环孢素A(CsA)处理急性单核细胞白血病细胞株THP-1后对柔红霉素(DNR)的耐药性及与多药耐药相关蛋白-1(MRP1)表达的关系,探讨CsA对THP-1细胞的化疗增敏作用及相关机制。方法:噻唑蓝(MTT)法、荧光分光光度法、共聚焦激光扫描显微镜(CLSM)法分别检测CsA处理前后50% THP-1细胞生长受抑的DNR浓度(IC50)、THP-1细胞内DNR含量及分布的变化。免疫细胞化学法、Western blot法检测CsA处理前后THP-1细胞MRP1蛋白表达量的变化。结果:CsA处理后THP-1细胞的IC50明显下降(1.5364±0.1751 vs 0.5588±0.0547,P<0.01),细胞内DNR含量明显增加(21.40±1.71 vs 131.96±16.45,P<0.01),且胞浆内分布更加均匀;同时MRP1蛋白表达量增高(0.2665±0.0042 vs 0.3169±0.0062,P<0.01)。结论:CsA可增加THP-1细胞对DNR的敏感性,降低其耐药性,并提示CsA对THP-1细胞耐药性的改变不是通过降低其MRP1蛋白表达量来实现的。
OBJECTIVE: To investigate the relationship between the drug resistance of daunorubicin (DNR) and the expression of multidrug resistance-associated protein-1 (MRP1) after treatment with CsA in THP-1 cells To investigate the chemosensitizing effect of CsA on THP-1 cells and its related mechanisms. Methods: The DNR concentration (IC50), the DNR of THP-1 cells in 50% THP-1 cells before and after CsA treatment were detected by MTT assay, fluorescence spectrophotometry and confocal laser scanning microscopy (CLSM) Changes in content and distribution. Immunocytochemistry and Western blot were used to detect the changes of MRP1 protein expression in THP-1 cells before and after CsA treatment. Results: The IC50 of THP-1 cells was significantly decreased after CsA treatment (1.5364 ± 0.1751 vs 0.5588 ± 0.0547, P <0.01), and the intracellular DNR content increased significantly (21.40 ± 1.71 vs 131.96 ± 16.45, P <0.01) The distribution of MRP1 protein increased (0.2665 ± 0.0042 vs 0.3169 ± 0.0062, P <0.01). Conclusion: CsA can increase the sensitivity of THP-1 cells to DNR and decrease the resistance of THP-1 cells to THP-1 cells, suggesting that the changes of drug resistance of THP-1 cells are not caused by the decrease of MRP1 protein expression.