| Objective:To preliminarily explore the role of astragaloside in reversing multi-drug resistance of gastric cancer cell lines SGC7901/DDP in vitro and the possible mechanisms.Methods:1.Drug-resistance of SGC7901/DDP was evaluated by MTT method;2.MTT assay was used to determine the cytotoxicity of astragaloside and verapamil,and the non-toxic concentrations of the two drugs were selected for further experiment;3.With reversal agent verapamil as positive control,MTT method was adopted to identify the role of non-toxic concentration of astragalosidein reversing multi-drug resistance of gastric cancer cell lines SGC7901/DDP;4.RT-PCR method was used to detect the effect of astragalosideon the expression of MDR1 gene;5.Western boltmethod was used to observe the effect of astragaloside on the expression of MDR1 gene encoding product P-glycoprotein.Results:1.The sensitivity of SGC7901/DDP cells to cisplatin was significantly weaker than SGC7901 cells,and the relative resistance index was 3.26,indicating certain drug resistance of this cell.2.Astragaloside can inhibit the proliferation of SGC7901/DDP drug-resistant cells,and was in a dose-dependent and dose-dependent relationship.Vallapamil had a strong inhibitory effect on drug-resistant cells,and the cumulative toxicity was significant.0.02,0.04,0.08 mg/mL Astragaloside and 10μg/mL verapamil on the SGC7901/DDP cells were not more than 10%inhibition rate,can be used as non-toxic concentration.3.MTT results showed that astragalosidecouldenhance the sensitivity of SGC7901/DDP cells to cisplatin so as to achieve the reverse of multidrug resistance,with concentration dependence.After treated with 0.02,0.04,and 0.08 mg/mL of astragaloside for 24 h,cells inhibition rates were 11.07±1.09,16.29±1.32 and 24.63±1.68,respectively,which were all higher than thoseof the control group 8.97±1.07;relative reverse times were 1.23,1.82 and 2.75,and relative reverse index of 0.01 mg/mL verapamil was 4.67.4.RT-PCR results showed that MDR1 mRNA level ofSGC7901/DDP cells treated with different concentrations of astragaloside was decreased.5.Western bolt results showed that relative expression of P-gp was gradually decreased with the increase of the concentration of astragaloside,and the P-gpwassignificantly reduced by 0.08 mg/ml of astragaloside.Conclusion:This experiment proves that non-toxic concentrationof astragaloside can reverse the multi-drug resistance of SGC7901/DDP cells in vitro,and its mechanism may be related to down-regulated expression of MDR1 gene and P-gp protein.0.08 mg/mL astragaloside has good ability to reverse the drug-resistance,although with a certain distance compared with 0.01 mg/mL verapamil,its side effectis significantly lower than the latter,so it can be used as an effective reverse regulator used in P-gp mediated multi-drug resistance of gastric cancer. |
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