As_2O_3对人胃癌多药耐药细胞模型的逆转耐药作用

THE EFFECT OF ARSENIC TRIOXIDE ON RESISTANCE REVERSAL IN CELL MODEL OF MULTIDRUG-RESISTANT GASTRIC CARCINOMA

目的体外建立人胃癌细胞多药耐药模型,研究多药耐药机制及三氧化二砷（As2O3）对其逆转耐药作用。方法采用阿霉素（ADM）浓度梯度递增法诱导建立人胃癌多药耐药细胞株SGC7901/ADM,MTT法检测SGC7901/ADM细胞对ADM、长春新碱（VCR）、紫杉醇（TAX）的敏感性及As2O3对其逆转耐药倍数,免疫细胞化学法（PV法）检测细胞P-糖蛋白（P-gp）、Caspase3表达,流式细胞仪检测细胞凋亡率。结果 SGC7901/ADM细胞对ADM、VCR和TAX均耐药,耐药倍数分别是7.49、7.56及5.33。在As2O3作用48h后,3种抗癌药物对SGC7901/ADM细胞生长抑制50%时的浓度（IC50）明显降低,差异有显著性（F=169.766-1 199.812,q=6.86-70.40,P〈0.01）;耐药细胞经As2O3及环孢素A（CsA）作用后,P-gp表达明显下调,差异有显著性（F=42.870,q=7.70-21.50,P〈0.01）;As2O3高浓度组细胞中Caspase3表达上调,差异有显著性（F=7.220,q=6.63,P〈0.05）。As2O3作用组SGC7901/ADM细胞凋亡率随As2O3浓度增加而增加（F=1 986.63,q=21.23-95.08,P〈0.05）。结论 SGC7901/ADM细胞具有多药耐药性,As2O3可逆转其耐药,并呈剂量依赖性。

Objective To establish A multidrug-resistant（MDR）cell model of gastric cancer in vitro,and study the effect of arsenic trioxide（As2O3）on MDR reversal. Methods MDR human gastric carcinoma cell line（SGC7901/ADM）was created in vitro by gradually increasing the density of adriamycin（ADM）.MTT assay was used to determine the sensitivity of SGC7901/ADM to ADM,VCR and TAX,and the drug resistance reversal fold（DRRF）of As2O3 were determined by MTT method;PV-method was used to detect the expressions of P-gp and Caspase3;flow cytometry（FCM）was applied to check apoptosis rate in tumor cells. Results SGC7901/ADM cells were resistant to ADM,VCR and TAX with a drug resistance fold of 7.49,7.56 and 5.33,respectively.After intervention with As2O3 for 48hours,the IC50 for the three anti-cancer drugs significantly reduced in SGC7901/ADM（F=169.766-1 199.812,q=6.86-70.40,P〈0.01）;P-gp expression levels（OD value）was significantly lower in As2O3 and CsA groups（F=42.870,q=7.70-21.50,P〈0.01）;Caspase3increased in As2O30.5μmoL/L group（F=7.220,q=6.63,P〈0.05）.Apoptosis rate was positively correlated with drug concentration and time（F=1 986.63,q=21.23-95.08,P〈0.05）. Conclusion SGC7901/ADM cell lines show multi-drug resistance,As2O3 can reverse its drug resistance manifesting dose-dependent.