ATP竞争性GSK-3抑制剂对人食管鳞癌细胞多药耐药的影响

目的: 研究ATP竞争性糖原合成酶激酶3(GSK-3)抑制剂6-溴靛玉红-3'-肟(BIO)作用于食管鳞癌细胞后, 对ATP结合盒(ABC)转运蛋白——P-糖蛋白(P-gp)和多药耐药相关蛋白2(MRP2)、凋亡抑制蛋白Bcl-2以及β-catenin表达的影响,探讨它们表达的相互关系,以及对细胞内游离ATP水平和P-gp、MRP2转运功能的影响,进而初步探讨GSK-3抑制剂对食管鳞癌细胞多药耐药的影响。方法: BIO作用于食管鳞癌EC-109细胞后,运用免疫荧光细胞化学法、流式细胞术以及ATP检测试剂盒分别检测细胞内 MRP2、P-gp、β-catenin和Bcl-2表达的改变、ABC转运蛋白的转运功能以及细胞内游离ATP水平的改变。结果: BIO作用食管鳞癌EC-109细胞后,加药组与对照组相比,β-catenin和Bcl-2在胞浆中的表达增强,并且β-catenin出现胞核内累积,MRP2在胞浆和胞膜中表达增强,P-gp在胞浆和胞膜中表达减弱;细胞内游离ATP水平增加;ABC转运蛋白的转运功能增强。结论: BIO处理食管鳞癌细胞后增强了细胞的多药耐药。

Effects of ATP-competitive GSK-3 inhibitor on multidrug resistance of human esophageal squamous-cell carcinoma cells

AIM: To study the changes and correlations of β-catenin,multidrug resistance-associated protein 2(MRP2),P-glycoprotein(P-gp),Bcl-2 and the free ATP level in esophageal squamous-cell carcinoma(ESCC) cell line EC-109 induced by ATP-competitive glycogen synthase kinase(GSK-3) inhibitor 6-bromoindirubin-3’-oxime(BIO).METHODS: The methods of flow cytometry,immunofluorescence,cytochemistry and ATP assay were used to study the expression levels of MRP2,P-gp,β-catenin and Bcl-2,the efflux capability of ATP-binding cassette(ABC) transporters,and the free ATP level in ESCC cells.RESULTS: After induced by BIO,up-regulation of β-catenin and Bcl-2 expression in cytoplasm and accumulation of β-catenin in nucleus were found in ESCC cells.The expression of MRP2 was up-regulated in cytoplasm and cell membrane.On the contrary,the expression of P-gp was down-regulated in cytoplasm and cell membrane.The free ATP level and the efflux capability of ABC transporters increased in ESCC cells.CONCLUSION: The multidrug resistance of ESCC cells in enhanced by BIO treatment.