Hepatitis C virus core protein stimulates hepatocyte growth: Correlation with upregulation of wnt-1 expression
Hepatitis C virus core protein stimulates hepatocyte growth: Correlation with upregulation of wnt-1 expression
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DOI：
10.1002/hep.20668
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发表时间：
2005-05-01
期刊：
影响因子：
13.5
通讯作者：
Li, JS
中科院分区：
文献类型：
--
作者：
Fukutomi, T;Zhou, YH;Li, JS
丙型肝炎病毒核心蛋白与人肝细胞癌的发生发展密切相关。在这里，我们报告了通过瞬时转染法表达丙型肝炎病毒核心蛋白，促进了人肝癌细胞系Huh-7细胞的增殖、DNA合成和细胞周期进展。转基因细胞的培养上清液也具有促进生长的作用。此外，一个全长的丙型肝炎病毒复制子，而不是一个没有核心基因的亚基因组复制子，显著刺激了瞬时转染的Huh-7.5细胞的生长。然而，二次转染亚基因组复制子Huh-7.5细胞的生长可以被核心基因所刺激，但不能被全长复制子中的其他结构基因所刺激。微阵列分析显示，在表达Huh-7细胞的核心蛋白中，已知的12,500个人类基因中有372个发生了三倍或更多的转录变化，大多数参与细胞生长或致癌信号的基因上调而不是下调。特别令人感兴趣的是WNT-1及其下游靶基因WISP-2的显著上调。事实上，针对WNT-1的小干扰RNA减弱了核心基因对细胞生长的刺激，而转导WNT-1基因的Huh-7细胞足以促进细胞增殖。与WNT-1蛋白的分泌一致，WNT-1转基因细胞的条件培养液促进了细胞的生长。总之，丙型肝炎病毒核心蛋白诱导Huh-7细胞增殖，无论是单独的还是在丙型肝炎病毒复制的背景下，这至少部分是通过转录上调与生长相关的基因，特别是WNT-1。
Hepatitis C virus (HCV) core protein has been implicated in the development of human hepatocellular carcinoma (HCC). Here we report that expression of HCV core protein by transient transfection increased cell proliferation, DNA synthesis, and cell cycle progression in Huh-7 cells, a human HCC-derived cell line. Culture supernatant from transfected cells also harbored a growth-promoting effect. Moreover, a full-length HCV replicon, but not a subgenomic replicon devoid of the core gene, significantly stimulated growth of transiently transfected Huh-7.5 cells. However, growth of the subgenomic replicon-containing Huh-7.5 cells could be stimulated by secondary transfection with core gene but not other structural genes present in the full-length replicon. Microarray analysis revealed threefold or more transcriptional changes in 372 of 12,500 known human genes in core protein expressing Huh-7 cells, with most genes involved in cell growth or oncogenic signaling, being upregulated rather than downregulated. Of particular interest is the marked upregulation of both wnt-1 and its downstream target gene WISP-2. Indeed, small interfering RNA against wnt-1 blunted growth stimulation by core gene, whereas transfection of Huh-7 cells with the wnt-1 gene sufficed to promote cell proliferation. Consistent with secretion of the wnt-1 protein, conditioned medium from wnt-1 transfected cells accelerated cell growth. In conclusion, HCV core protein induces Huh-7 cell proliferation whether alone or in the context of HCV replication, which is at least partly mediated by transcriptional upregulation of growth-related genes, in particular wnt-1.