HBV感染与宿主细胞高尔基体α^1甘露糖苷酶Ⅰ活性关系的研究

Research on the relationship between HBV infection and activity of Golgi MⅠ

目的:研究慢性HBV感染与宿主细胞高尔基体α1甘露糖苷酶I(Golgi MⅠ)活性的关系。方法:培养Hep G2细胞和Hep G2.2.15细胞,采用蔗糖密度梯度离心法分离纯化高尔基复合体,比色法检测Golgi MⅠ的活性,RT-PCR法检测基因MAN1A1(人甘露糖苷酶1A类成员1)、MAN1A2和MAN1C1 m RNA的表达,Western blot法检测上述3种基因编码蛋白的表达情况。结果:与Hep G2细胞相比,Hep G2.2.15细胞Golgi MⅠ的活性明显升高,MAN1A1、MAN1A2的m RNA水平显著上调,所编码的MAN1A1、MAN1A2蛋白量也显著增加,而MAN1C1的m RNA和蛋白的表达量无明显变化。结论:HBV感染可提高宿主细胞Golgi MⅠ的活性,这一作用可能是通过上调MAN1A1、MAN1A2基因m RNA和蛋白表达的量介导。

Objective：To investigate whether HBV infection would increase the activity of Golgi MⅠin Hep G2 cell line. Methods：Hep G2 cells and Hep G2.2.15 cells were cultured,and Golgi complexes of these cells were purified by sucrose density gradient centrifugation.The activity of Golgi MⅠin Golgi complexes was detected by colorimetric method. The m RNA expression of MAN1A1、MAN1A2 and MAN1C1 was performed by RT-PCR method. The protein expression of MAN1A1、MAN1A2 and MAN1C1 was performed by Western blot method. Results：It was found that the activity of Golgi MⅠincreased,the m RNA and protein expression of MAN1A1 and MAN1A2 increased too in Hep G2.2.15 cells compared with Hep G2 cells,but there was no difference in the m RNA and protein expression of MAN1C1 between two groups. Conclusion：HBV infection could increase the activity of Golgi MⅠ by up-regulating the m RNA and protein expression of MAN1A1 and MAN1A2.