Prominent steatosis with hypermetabolism of the cell line permissive for years of infection with hepatitis C virus.
Prominent steatosis with hypermetabolism of the cell line permissive for years of infection with hepatitis C virus.
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DOI：
10.1371/journal.pone.0094460
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Kanai T
中科院分区：
文献类型：
--
作者：
Sugiyama K;Ebinuma H;Nakamoto N;Sakasegawa N;Murakami Y;Chu PS;Usui S;Ishibashi Y;Wakayama Y;Taniki N;Murata H;Saito Y;Fukasawa M;Saito K;Yamagishi Y;Wakita T;Takaku H;Hibi T;Saito H;Kanai T
大多数HCV感染的实验都是使用裂解感染系统进行的，其中HCV感染的细胞不可避免地死亡。在这里，为了阐明HCV感染细胞在更稳定条件下的代谢改变，我们建立了HCV持续感染细胞系，命名为HPI细胞。该细胞系显示出显著的脂肪变性，并支持HCV感染超过2年，这是有史以来报道的最长时间。它使我们能够分析代谢在HCV感染的细胞整合代谢组学和表达阵列。结果表明，随着胆固醇、链甾醇（胆固醇前体）和脂肪酸库的实际增加，胆固醇和脂肪酸生物合成的限速酶上调。值得注意的是，戊糖磷酸途径促进了葡萄糖-6-磷酸脱氢酶，一种视网膜限制酶，与NADPH的实际增加的显着上调。在其下游，嘌呤合成酶也被上调，导致嘌呤增加。与常见癌症相反，TCA循环比糖酵解途径优先促进，大多数氨基酸显著增加。有趣的是，一些基因控制的核因子（红细胞衍生2）样2（Nrf 2），抗氧化和代谢的主调节器，组成性上调HPI细胞。敲低Nrf 2基因可显著降低脂肪变性和HCV感染，表明Nrf 2及其靶基因在代谢改变和HCV感染中起重要作用。结论：HPI细胞是一个真正的HCV持续感染细胞系，支持HCV感染多年。该细胞系在高代谢状态下持续显著的脂肪变性，产生各种代谢产物。因此，HPI细胞是一种有效的研究工具，不仅对持续性HCV感染，而且对肝脏代谢，克服裂解感染系统的缺点。
Most of experiments for HCV infection have been done using lytic infection systems, in which HCV-infected cells inevitably die. Here, to elucidate metabolic alteration in HCV-infected cells in a more stable condition, we established an HCV-persistently-infected cell line, designated as HPI cells. This cell line has displayed prominent steatosis and supported HCV infection for more than 2 years, which is the longest ever reported. It enabled us to analyze metabolism in the HCV-infected cells integrally combining metabolomics and expression arrays. It revealed that rate-limiting enzymes for biosynthesis of cholesterol and fatty acids were up-regulated with actual increase in cholesterol, desmosterol (cholesterol precursor) and pool of fatty acids. Notably, the pentose phosphate pathway was facilitated with marked up-regulation of glucose-6-phosphate dehydrogenase, a rete-limiting enzyme, with actual increase in NADPH. In its downstream, enzymes for purine synthesis were also up-regulated resulting in increase of purine. Contrary to common cancers, the TCA cycle was preferentially facilitated comparing to glycolysis pathway with a marked increase of most of amino acids. Interestingly, some genes controlled by nuclear factor (erythroid-derived 2)-like 2 (Nrf2), a master regulator of antioxidation and metabolism, were constitutively up-regulated in HPI cells. Knockdown of Nrf2 markedly reduced steatosis and HCV infection, indicating that Nrf2 and its target genes play important roles in metabolic alteration and HCV infection. In conclusion, HPI cell is a bona fide HCV-persistently-infected cell line supporting HCV infection for years. This cell line sustained prominent steatosis in a hypermetabolic status producing various metabolites. Therefore, HPI cell is a potent research tool not only for persistent HCV infection but also for liver metabolism, overcoming drawbacks of the lytic infection systems.
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