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Objective: Determination of aloe emodin (Aloe Emodin, AE) cytotoxicity of human lung adenocarcinoma cell line A549 and A549/DDP determine aloe-emodin the cytotoxic concentration; detect this concentration of aloe emodin can reverse the A549 / DDP cell lines to cisplatin resistance, and to explore its possible mechanism. Method: 1, take the logarithmic growth phase A549, the of A549/DDP, establishment of aloe emodin group: the concentration of 2.5μg · mL -1 , 5 μg · mL -1 solvent control, 10μg · mL -1 , 15μg · mL -1 , 20μg · mL -1 ; negative control group: plus culture medium; Group: 0.1% dimethyl (Diaminobenzidine, DMSO) and blank control group: without cells. Aloe emodin MTT assay the A549 and A549/DDP of cells and 24 h, 48 h, 72 h cell toxicity, determine the aloe emodin 48 h non-cytotoxic concentrations. 2 cisplatin (concentration: 0.125μg · mL -1 , 0.25μg · mL -1 0.5μg · mL -1 , 1.0μg · mL -1 , 2.0μg · mL -1 , 4.0μg · mL -1 , 8.0μg · mL -1 , 16μg · mL -1 , 24μg · mL -1 , 32μg · mL -1 ) dealt with separately in A549 of A549/DDP 48 h, MTT assay inhibitory effect, and obtained A549/DDP multiples to DDP resistance. 3, MTT assay non-cytotoxic concentration of aloe emodin and cisplatin (concentration: 0.125μg · mL -1 0.25μg · mL -1 0.5μg · mL -1 , 1.0μg · mL -1 , 2.0μg · mL -1 , 4.0μg · mL -1 , 8.0μg · mL -1 , 16μg · mL -1 , 24μg · mL -1 , 32μg · mL - 1 ) MS inhibitory effect of A549/DDP, aloe-emodin A549/DDP resistance reversal multiples is obtained. 4, non-cytotoxic concentration of aloe emodin A549/DDP strains of cell growth curve and doubling time of the impact. 5, inductively coupled plasma mass spectrometry (ICP-MS) method A549 and A549/DDP cells cisplatin concentration discussed aloe emodin reversal of multidrug resistance mechanism. Results: 1 MTT colorimetric results show: AE can significantly inhibit the proliferation of the A549 the strains and A549/DDP strain cells cultured in vitro, and the degree of inhibition of increase in with aloe emodin time and increase in the concentration of and was a certain time, dose (2.5-20μg · mL -1 )-dependent the AE itself cytotoxicity, can inhibit the proliferation of lung adenocarcinoma cell; less than or equal to 5μg · mL -1 < / sup> AE for 48 h the of A549 and A549/DDP two cells had no significant toxicity (IR <10%), the 5μg · mL -1 non-cytotoxic concentration for AE. A549 cells IC 50 1.344μg · mL -1 of A549/DDP cells IC 50 of as 16.81μg · mL -1 , A549/DDP resistant multiple of 12.51 times; cytotoxic concentration (5 μg · mL -1 ) the AE and DDP combination of A549/DDP cells IC 50 reduced 5.86μg · mL -1 the reversal fold to 2.87, indicating that the AE enhanced the killing effect of the DDP on A549/DDP cell lines, that when combined with DDP can restore the DDP on cells sensitivity A549/DDP, show that the the AE has chemosensitizing role growth curve consistent with the results and the cell count. 3, inductively coupled plasma mass spectrometry results: cytotoxic concentration (5 μg · mL -1 ) the AE and 4μg of · mL -1 DDP combination, can significantly increase the A549 / DDP strain DDP intracellular accumulation, while no significant accumulation of DDP increased in A549 cells. Conclusion: AE on the growth of A549 the strains and A549/DDP cell lines significantly inhibited, and increases with the increase of the AE concentration and duration of action and growth inhibition rate was drug concentration - time-dependent manner. Show that AE two cell proliferation inhibition. The 2,5 μg · mL -1 for AE cytotoxic concentration. The concentration AE can significantly enhance DDP inhibited proliferation of human lung adenocarcinoma cells resistant strains A549/DDP AE part of the role reversal A549/DDP cells resistant to cisplatin. 3,5 μg · mL -1 AE significantly increased DDP content A549/DDP cells, without significant increase in the concentration of A549 cells DDP, shows that the role of the reversal of drug resistance, and increased intracellular drug concentration.
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