A new hepatoma cell line exhibiting high susceptibility to hepatitis B virus infection
A new hepatoma cell line exhibiting high susceptibility to hepatitis B virus infection
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DOI：
10.1016/j.bbrc.2019.05.126
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发表时间：
2019-06-12
影响因子：
3.1
通讯作者：
Kato，Nobuyuki
中科院分区：
文献类型：
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作者：
Ueda，Youki;Gu，Weilin;Kato，Nobuyuki
B型肝炎病毒（HBV）感染增加了肝硬化和肝细胞癌的风险，需要终身治疗，已成为一个主要的全球健康问题。然而，HBV的生命周期所必需的宿主因素仍不清楚，需要开发新的药物，来自人肝癌细胞系HepG 2的细胞被工程改造为过表达牛磺胆酸钠共转运多肽（NTCP：HBV的受体），称为HepG 2/NTCP细胞，被广泛用作基于细胞的HBV感染和复制系统，用于HBV的研究。我们最近发现过表达NTCP的人肝癌细胞系Li 23衍生细胞（从Li 23细胞亚克隆的A8细胞），其基因表达谱与HepG 2/NTCP细胞不同，也对HBV感染敏感。然而，A8细胞的HBV易感性约为HepG 2/NTCP细胞的1/100。由于我们认为客观评价抗HBV试剂需要多细胞检测系统，正如我们以前在丙型肝炎病毒研究中所证明的那样，我们在这里试图开发一种新的Li 23细胞衍生的检测系统，其等同于使用HepG 2/NTCP细胞的检测系统。通过对A8细胞的重复亚克隆，我们成功地建立了一个新的细胞系（A8.15.78.10），其表现出与HepG 2/NTCP细胞相当的高HBV易感性。对A8.15.78.10细胞的表征显示，HBV易感性的增加与NTCP的蛋白质和糖基化水平的增加以及STING表达的减少相关，STING是一种促进先天免疫的因素。最后，我们使用A8.15.78.10细胞或HepG 2/NTCP细胞通过HBV/secNL报告基因测定对HBV进入抑制剂（环孢菌素A和罗格列酮）进行了比较评估。结果证实，环孢菌素A在两种细胞系中均表现出抗HBV活性，如先前所报道的。然而，我们发现罗格列酮在A8.15.78.10细胞中没有显示出抗HBV活性，尽管它在HepG 2/NTCP细胞中起作用，如先前报道的。这表明环孢菌素A和罗格列酮之间抗HBV活性的差异是由于用于测定的细胞类型不同。总之，需要使用不同类型细胞的多个检测系统来客观和公正地评价抗HBV试剂。
Hepatitis B virus (HBV) infection, which increases the risk of cirrhosis and hepatocellular carcinoma and requires lifelong treatment, has become a major global health problem. However, host factors essential to the HBV life cycle are still unclear, and the development of new drugs is needed.Cells derived from the human hepatoma cell line HepG2 and engineered to overexpress sodium taurocholate cotransporting polypeptide (NTCP: a receptor for HBV), termed HepG2/NTCP cells, are widely used as the cell-based HBV infection and replication systems for HBV research. We recently found that human hepatoma cell line Li23-derived cells overexpressing NTCP (A8 cells subcloned from Li23 cells), whose gene expression profile was distinct from that of HepG2/NTCP cells, were also sensitive to HBV infection. However, the HBV susceptibility of A8 cells was around 1/100 that of HepG2/NTCP cells. Since we considered that plural cell assay systems will be needed for the objective evaluation of anti-HBV reagents, as we previously demonstrated in hepatitis C virus research, we here attempted to develop a new Li23 cell-derived assay system equivalent to that using HepG2/NTCP cells. By repeated subcloning of A8 cells, we successfully established a new cell line (A8.15.78.10) exhibiting high HBV susceptibility equal to that of HepG2/NTCP cells. Characterization of A8.15.78.10 cells revealed that the increase of HBV susceptibility was correlated with increases in the protein and glycosylation levels of NTCP, and with decreased expression of STING, a factor contributing to innate immunity. Finally, we performed a comparative evaluation of HBV entry inhibitors (cyclosporin A and rosiglitazone) by an HBV/secNL reporter assay using A8.15.78.10 cells or HepG2/NTCP cells. The results confirmed that cyclosporin A exhibited anti-HBV activity in both cell lines, as previously reported. However, we found that rosiglitazone did not show the anti-HBV activity in A8.15.78.10 cells, although it worked in HepG2/NTCP cells as previously reported. This suggested that the difference in anti-HBV activity between cyclosporin A and rosiglitazone was due to the different types of cells used for the assay. In conclusion, plural assay systems using different types of cells are required for the objective and impartial evaluation of anti-HBV reagents.