人胆囊癌细胞系GBC-SD的多药耐药机制

Mechanisms of multidrug resistance in human gallbladder carcinoma cell line GBC-SD

目的 探讨人胆囊癌细胞系GBC SD耐药性产生的机制 ,建立先天性耐药的人胆囊癌细胞模型。方法 用逆转录 聚合酶链反应 (RT PCR )检测GBC SD细胞多药耐药相关蛋白(MDR1)、多药耐药相关蛋白 (MRP)和肺耐药蛋白 (LRP)基因扩增 ;免疫细胞化学染色检测该细胞系P 糖蛋白 (P gp)、MRP和LRP蛋白表达 ;流式细胞术测定P gp、谷胱甘肽 S 转移酶 (GST π)和DNA拓扑异构酶Ⅱ (TOPOⅡ )表达水平 ;用四甲基偶氮唑蓝 (MTT)比色法检测该细胞对 8种抗癌药物的敏感性。结果 人胆囊癌GBC SD细胞中MDR1、MRP和LRP基因呈阳性扩增 ,P gp、MRP、LRP蛋白表达阳性 ;GST π和TOPOⅡ的表达水平与对照组细胞差异无显著性。四唑盐比色结果显示 ,GBC SD细胞对 8种抗肿瘤药物均有程度不同的耐受性 ,耐药倍数从 5 .3倍到 42倍不等。结论 人胆囊癌细胞系GBC SD是一株先天性耐药的恶性肿瘤细胞系 ,其耐药性的产生与MDR1、MRP、LRP基因扩增有关。

Objective To study the mechanisms of multidrug resistance (MDR) in human gallbladder carcinoma cell line and establish the MDR cell model of human gallbladder carcinoma.Methods The amplification of MDR genes (MDR1,MRP and LRP) in human gallbladder carcinoma cell line BGC-SD was detected by using reverse transcript polymerase chain reaction (RT-PCR).The immunohistochemistry was performed to detect the expression levels of P-gp,MRP and LRP.The flow cytometry (FCM) was carried out to determine the expression levels of P-gp,GST-π and TOPOII.The IC50 of 8 anticancer drugs to GBC-SD cells was determined by MTT method.Results The amplification of MDR genes (MDR1,MRP,LRP) and the expression of P-gp,MRP and LRP were positive in GBC-SD cells.There was no significant difference in the expression levels of GST-π and TOPOII between the GBC-SD cells and control group.GBC-SD cells were less sensitive to 8 anticancer drugs.Conclusion With the amplification of the genes MDR1,MRP and LRP,GBC-SD is an intrinsic MDR cell line.It can be used as an optimal MDR cell model for further study of multidrug resistance of gallbladder carcinoma.