HCV感染相关的肝细胞性肝癌中精氨酸酶Ⅱ的表达

Expression of Arginase 2 in Hepatitis C Virus related Hepatocellular Carcinoma

目的通过对丙型肝炎病毒-原发性肝细胞肝癌（HCV-HCC）组织及细胞中精氨酸酶Ⅱ（Arg-Ⅱ）的表达进行分析，并以RNA干扰技术阻断其在细胞内的表达，初步研究Arg-Ⅱ表达在HCV-HCC发病中的意义。方法首先以基因转染技术分别将HCV基因组和对照质粒导入HuH7细胞中，以逆转录-实时荧光PCR检测HCV基因组的转录并作细胞生长曲线测定，以Western blotting技术研究HCV基因组转染对细胞内Arg-Ⅱ表达的影响，以免疫组化技术分析HCV-HCC组织中Arg-Ⅱ的表达水平，运用RNA干扰技术对Arg-Ⅱ作初步的功能分析。结果以基因转染技术分别将HCV基因组和对照质粒导入HuH7，成功建立了HuH7-HCV和HuH7-vector两个细胞系，逆转录-实时荧光PCR检测表明HuH7-HCV细胞的HCV转录水平为0．5～5拷贝／细胞。Western blotting结果显示，与对照细胞相比，HuH-7-HCV细胞的Arg-Ⅱ含量升高3．9倍。免疫组化技术分析HCV-HCC组织标本中Arg-Ⅱ的表达，结果呈强阳性，而对照标本中基本不表达。采用RNA干扰方法阻断HuH7-HCV细胞的Arg-Ⅱ表达，发现细胞增殖被明显抑制。结论Arg-Ⅱ在HCV阳性肝细胞系中及HCV-HCC组织中异常表达并可能参与HCV-HCC的发病机制。

To study expression of Arginase Ⅱ in HCV positive cells and in HCV-HCC tissues and evaluate its significance in HCV-HCC development. Methods HuH7 cells were stably transfected with HCV genome. HCV genome transcription was determined by RT-PCR, and cell growth curve was measured. Western blotting and immunohistochemistry were used to study the expression level of Arginase Ⅱ in HCV positive HuH7 cells and in HCV-HCC tissues. SiRNA was performed to study the possible meaning of Arginase Ⅱ expression to the cell growth. Results HuH7-HCV and HuH7-vector cell lines were successfully established. In HuH7-HCV, HCV transcription level was 0.5 - 5 copy/cell. Arginase Ⅱ was over-expressed in HCV positive HuH7 cells and HCV-HCC tissues, which was 3.9 times higher in HuH7-HCV than control group. SiRNA of Arginase Ⅱ inhibited cell growth. Conclusion Arginase Ⅱ over-expression may play an important role in HCV-HCC development.