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Ezrin上调HepG2细胞膜转运蛋白MRP2的表达

, PP. 1951-1954

Keywords: Ezrin基因缺失,胆汁淤积,多药耐药相关蛋白

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Abstract:

目的建立Ezrin基因表达缺失的HepG2细胞株,并研究其在胆汁淤积时对HepG2细胞胆汁酸转运蛋白多药耐药相关蛋白2(multidrugresistance-associatedprotein2,MRP2)的影响。方法不同位点干扰的Ezrin-shRNA质粒转染HepG2细胞,筛选Ezrin基因表达缺失的HepG2细胞。分别提取细胞总蛋白和膜蛋白,检测Ezrin基因表达缺失是否影响HepG2细胞胆汁酸转运蛋白MRP2的表达。结果RT-qPCR结果证实,Ezrin-shRNA#3质粒转染HepG2细胞后成功抑制了Ezrin基因表达,抑制率为70%。蛋白免疫印迹表明,Ezrin基因表达缺失后,HepG2细胞胆汁酸转运蛋白MRP2表达增高,但与对照组相比,差异无统计学意义(P>0.05)。蛋白免疫共沉淀检测生物素化MRP2膜蛋白表明,Ezrin基因表达缺失后,无论是胞浆中还是细胞膜上,MRP2膜蛋白均表达增高(P<0.05)。结论Ezrin基因表达缺失后能够上调HepG2细胞中MRP2膜蛋白,使其表达增高。

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