Mitomycin, 5-fluorouracil, leflunomide, and mycophenolic acid directly promote hepatitis B virus replication and expression in vitro
Mitomycin, 5-fluorouracil, leflunomide, and mycophenolic acid directly promote hepatitis B virus replication and expression in vitro
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丝裂霉素、5-氟尿嘧啶、来氟米特、麦考酚酸直接促进乙型肝炎病毒体外复制和表达
DOI：
10.1186/s12985-020-01339-5
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发表时间：
2020-07
期刊：
影响因子：
4.8
通讯作者：
Sun Dianxing
中科院分区：
文献类型：
--
作者：
Ruan Jie;Sun Shuo;Cheng Xin;Han Pengyu;Zhang Yinge;Sun Dianxing
背景乙型肝炎病毒再激活是接受细胞毒化疗或免疫抑制治疗的乙型肝炎病毒（HBV）感染患者常见的并发症。这种临床现象不仅发生在明显的乙型肝炎病毒感染患者中，而且也发生在乙型肝炎病毒感染已治愈的患者中。先前的研究已证实表阿霉素和地塞米松可以直接刺激乙型肝炎病毒的复制和表达，而不是通过免疫抑制间接刺激乙型肝炎病毒的复制和表达。丝裂霉素和5-氟尿嘧啶目前用作癌症患者的细胞毒性化疗药物。来氟米特和麦考酚酸被认为是治疗自身免疫性疾病的免疫抑制剂，大量临床研究报道这些药物可以重新激活乙肝病毒复制。本研究旨在探讨丝裂霉素、5-氟尿嘧啶、来氟米特和麦考酚酸是否直接而非通过免疫抑制间接诱导HBV再激活。方法以HepG2.2.15和HBV-NLuc-35细胞为细胞模型，观察丝裂霉素、5-氟尿嘧啶、来氟米特和麦考酚酸对HBV复制和表达的影响。接下来，通过天然琼脂糖凝胶电泳（NAGE）、定量PCR（qPCR）、荧光素酶测定和HBV e抗原（HBeAg）酶联免疫吸附测定（ELISA），我们检测了这些药物诱导的HBV复制和表达的变化。我们还研究了拉米夫定是否可以抑制观察到的表型。采用SPSS 18.0软件进行统计分析，采用单因素方差分析进行多组比较。结果随着丝裂霉素、5-氟尿嘧啶、来氟米特、麦考酚酸浓度的增加，HepG2.2.15细胞中HBV衣壳和HBeAg的表达增加。这种现象也在 HBV-NLuc-35 细胞中得到证实，衣壳的表达和荧光素酶活性以相同的浓度依赖性方式增加。 HepG2.2.15细胞内衣壳DNA和胞外HBV DNA的复制水平逐渐增加，并呈剂量依赖性。此外，虽然表柔比星、丝裂霉素、5-氟尿嘧啶、地塞米松、来氟米特和麦考酚酸增强了HBV复制，但拉米夫定抑制了这一过程。结论我们的研究证实，丝裂霉素、5-氟尿嘧啶、来氟米特和麦考酚酸在体外直接上调HBV复制和表达。这种效应不仅在 HepG2.2.15 细胞中进行了研究，还在 HBV-NLuc-35 复制系统中进行了研究。此外，核苷类似物如拉米夫定 (LAM) 可以防止这种效应。因此，对于HBV感染的患者，在接受细胞毒化疗或免疫抑制治疗之前，有必要进行预防性抗病毒治疗。
BackgroundReactivation of hepatitis B virus is a common complication that occurs in patients with hepatitis B virus (HBV) infection who have received cytotoxic chemotherapy or immunosuppressive therapy. This clinical phenomenon not only occurs in overt HBV infection patients but also occurs in patients with resolved HBV infection. Previous research has confirmed that epirubicin and dexamethasone can stimulate HBV replication and expression directly rather than indirectly through immunosuppression. Mitomycin and 5-fluorouracil are currently used as cytotoxic chemotherapy drugs for cancer patients. Leflunomide and mycophenolic acid are regarded as immunosuppressants for autoimmune diseases, and numerous clinical studies have reported that these drugs can reactivate HBV replication. In this study, we aimed to investigate whether mitomycin, 5-fluorouracil, leflunomide and mycophenolic acid induce HBV reactivation directly rather than indirectly through immunosuppression.MethodsTo observe the effect of mitomycin, 5-fluorouracil, leflunomide and mycophenolic acid on HBV replication and expression, we employed HepG2.2.15 and HBV-NLuc-35 cells as a cell model. Next, by native agarose gel electrophoresis (NAGE), quantitative PCR (qPCR), luciferase assay and HBV e antigen (HBeAg) enzyme-linked immunosorbent assay (ELISA) we detected changes in HBV replication and expression induced by these drugs. We also investigated whether lamivudine could inhibit the observed phenotype. SPSS 18.0 software was employed for statistical analysis, One-way ANOVA was used to compare multiple groups.ResultsExpression of HBV capsids and HBeAg in HepG2.2.15 cells was increased by increasing concentration of mitomycin, 5-fluorouracil, leflunomide, and mycophenolic acid. This phenomenon was also demonstrated in HBV-NLuc-35 cells, and the expression of capsids and luciferase activity increased in the same concentration-dependent manner. Replication levels of intracellular capsid DNA and extracellular HBV DNA in HepG2.2.15 cells gradually increased in a dose-dependent manner. In addition, although epirubicin, mitomycin, 5-fluorouracil, dexamethasone, leflunomide and mycophenolic acid enhanced HBV replication, lamivudine inhibited this process.ConclusionOur study confirmed that mitomycin, 5-fluorouracil, leflunomide and mycophenolic acid directly upregulated HBV replication and expression in vitro. This effect was investigated not only in HepG2.2.15 cells but also in the HBV-NLuc-35 replication system. Moreover, this effect could be prevented by nucleoside analogs, such as lamivudine (LAM). Thus, for patients with HBV infection, prophylactic antiviral therapy is necessary before receiving cytotoxic chemotherapy or immunosuppressive therapy.
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影响因子：
5.3
作者：
通讯作者：
--
影响因子：
29.4
作者：
J. Wands;C. Chura;F. Roll;W. Maddrey
通讯作者：
J. Wands;C. Chura;F. Roll;W. Maddrey
通过重新设计的病毒聚合酶翻译，具有复制能力的传染性乙型肝炎病毒载体携带大量转基因。
影响因子：
3.7
作者：
Wang Z;Wu L;Cheng X;Liu S;Li B;Li H;Kang F;Wang J;Xia H;Ping C;Nassal M;Sun D
通讯作者：
Sun D
DOI：
10.1002/hep.27609
发表时间：
2015-02
期刊：
Hepatology (Baltimore, Md.)
影响因子：
--
作者：
Di Bisceglie AM;Lok AS;Martin P;Terrault N;Perrillo RP;Hoofnagle JH
通讯作者：
Hoofnagle JH
影响因子：
39.2
作者：
V. Lo Re;M. Schuster
通讯作者：
V. Lo Re;M. Schuster