Ezrin上调HepG2细胞膜转运蛋白MRP2的表达

发布时间：2018-06-02 12:39

本文选题：Ezrin基因缺失 + 胆汁淤积　；参考：《第三军医大学学报》2015年19期

【摘要】：目的建立Ezrin基因表达缺失的Hep G2细胞株,并研究其在胆汁淤积时对Hep G2细胞胆汁酸转运蛋白多药耐药相关蛋白2(multidrug resistance-associated protein 2,MRP2)的影响。方法不同位点干扰的Ezrin-shRNA质粒转染Hep G2细胞,筛选Ezrin基因表达缺失的Hep G2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Hep G2细胞胆汁酸转运蛋白MRP2的表达。结果 RT-q PCR结果证实,Ezrin-shRNA#3质粒转染Hep G2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,Hep G2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(P0.05)。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(P0.05)。结论 Ezrin基因表达缺失后能够上调Hep G2细胞中MRP2膜蛋白,使其表达增高。
[Abstract]:Objective to establish a Hep G2 cell line with missing Ezrin gene expression and to study its effect on the bile acid transporter multidrug resistance associated protein 2(multidrug resistance-associated protein 2 MRP2in Hep G2 cells during cholestasis. Methods Hep G2 cells were transfected with Ezrin-shRNA plasmid with different site interference. Hep G2 cells with missing Ezrin gene expression were screened. Total protein and membrane protein were extracted, and the expression of bile acid transporter (MRP2) in Hep G2 cells was detected by Ezrin gene deletion. Results the results of RT-q PCR confirmed that the expression of Ezrin gene was inhibited successfully by Ezrin-shRNA #3 plasmid transfection into Hep G2 cells, and the inhibition rate was 70%. Western blot showed that the expression of bile acid transporter MRP2 in Hep G2 cells was increased after the deletion of Ezrin gene expression, but there was no significant difference compared with the control group (P 0.05). The detection of biotinylated MRP2 membrane protein by immunoprecipitation showed that the expression of MRP2 membrane protein increased in both cytoplasm and cell membrane after the deletion of Ezrin gene. Conclusion Ezrin gene expression can up-regulate and increase the expression of MRP2 membrane protein in Hep G2 cells.
【作者单位】：第三军医大学西南医院全军消化病研究所;
【基金】：国家自然科学基金面上项目(81370560,81170430);国家自然科学基金青年科学基金项目(81100280)~~
【分类号】：R575.6

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