Comparison between cytomegalovirus promoter and elongation factor-1 alpha promoter-driven constructs in the establishment of cell lines expressing hepatitis C virus core protein
Comparison between cytomegalovirus promoter and elongation factor-1 alpha promoter-driven constructs in the establishment of cell lines expressing hepatitis C virus core protein
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DOI：
10.1016/s0166-0934(96)02143-x
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发表时间：
1997-02-01
影响因子：
3.1
通讯作者：
Wands, JR
中科院分区：
文献类型：
--
作者：
Tokushige, K;Moradpour, D;Wands, JR
建立稳定表达丙型肝炎病毒（HCV）核心蛋白的细胞系对研究丙型肝炎病毒的发病机制具有重要意义。利用巨细胞病毒（CMV）或延伸因子-1 α (EF-1 α)启动子驱动的表达载体，生成了表达HCV核心蛋白的人和小鼠细胞系。瞬时转染后，HCV核心蛋白在所有细胞系中均有表达。然而，稳定表达HCV核心蛋白的人肝细胞癌（HCC）和小鼠骨髓瘤细胞系仅使用EF-1 α启动子驱动的构建体建立。相比之下，使用CMV启动子驱动的表达载体，在这些细胞系中成功地获得了乙型肝炎病毒（HBV）中间包膜蛋白（MHBs）的稳定表达。利用嵌合MHBs/HCV核心蛋白构建物，发现HCV核心蛋白的前69个氨基酸对CMV启动子具有抑制活性。表达HCV核心蛋白的克隆细胞系的生长速度比不表达的细胞系慢。然而，在表达HCV核心蛋白的稳定细胞系中，未观察到形态学变化和细胞死亡。这些结果表明，HCV核心蛋白对HCC和骨髓瘤细胞系没有直接的细胞毒性，但需要特定的启动子元件来建立核衣壳结构蛋白的稳定表达。版权所有1997爱思唯尔科学有限公司
The establishment of stable cell lines expressing the hepatitis C virus (HCV) core protein may be important for studies of HCV pathogenesis. Human and mouse cell lines were generated expressing the HCV core protein using expression vectors driven by either the cytomegalovirus (CMV) or elongation factor-1 alpha (EF-1 alpha) promoters. Following transient transfection, HCV core protein was expressed in all cell lines. However, stable human hepatocellular carcinoma (HCC) and murine myeloma cell lines expressing the HCV core protein were only established using constructs driven by the EF-1 alpha promoter. In contrast, stable expression of the hepatitis B virus (HBV) middle envelope protein (MHBs) was obtained successfully in these cell lines using an expression vector driven by the CMV promoter. Inhibitory activity of the first 69 amino acids of the HCV core protein on the CMV promoter was found by using chimeric MHBs/HCV core protein constructs. Growth of cloned cell lines expressing the HCV core protein was slower than that of nonexpressing cell lines. However, morphological changes and cell death were not observed in the stable cell lines expressing HCV core protein. These results indicate that the HCV core protein was not directly cytotoxic to HCC and myeloma cell lines but that specific promoter elements are required to establish stable expression of the nucleocapsid structural protein. Copyright (C) 1997 Elsevier Science B.V.