Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium.
Stem Cell Biology and Embryology, University of Leuven, O&N IV Herestraat 49 - bus 804, 3000 Leuven, Belgium.
Biochem Pharmacol. 2015 Aug 1;96(3):190-201. doi: 10.1016/j.bcp.2015.06.003. Epub 2015 Jun 9.
A number of statins, the cholesterol-lowering drugs, inhibit the in vitro replication of hepatitis C virus (HCV). In HCV-infected patients, addition of statins to the earlier standard of care therapy (pegIFN-α and ribavirin) resulted in increased sustained virological response rates. The mechanism by which statins inhibit HCV replication has not yet been elucidated. In an attempt to gain insight in the underlying mechanism, hepatoma cells carrying an HCV replicon were passaged in the presence of increasing concentrations of fluvastatin. Fluvastatin-resistant replicon containing cells could be generated and proved ∼8-fold less susceptible to fluvastatin than wild-type cultures. The growth efficiency of the resistant replicon containing cells was comparable to that of wild-type replicon cells. The fluvastatin-resistant phenotype was not conferred by mutations in the viral genome but is caused by cellular changes. The resistant cell line had a markedly increased HMG-CoA reductase expression upon statin treatment. Furthermore, the expression of the efflux transporter P-gp was increased in fluvastatin-resistant replicon cells (determined by qRT-PCR and flow cytometry). This increased expression resulted also in an increased functional transport activity as measured by the P-gp mediated efflux of calcein AM. In conclusion, we demonstrate that statin resistance in HCV replicon containing hepatoma cells is conferred by changes in the cellular environment.
一些他汀类药物,即降低胆固醇的药物,可抑制丙型肝炎病毒(HCV)的体外复制。在 HCV 感染患者中,他汀类药物联合早期标准护理疗法(聚乙二醇干扰素-α和利巴韦林)可提高持续病毒学应答率。他汀类药物抑制 HCV 复制的机制尚未阐明。为了深入了解潜在的机制,在存在逐渐增加的氟伐他汀浓度的情况下,将携带 HCV 复制子的肝癌细胞进行传代。可以产生对氟伐他汀具有耐药性的复制子含有细胞,并且比野生型培养物对氟伐他汀的敏感性降低约 8 倍。耐药性复制子含有细胞的生长效率与野生型复制子细胞相当。耐药表型不是由病毒基因组中的突变引起的,而是由细胞变化引起的。在用他汀类药物处理时,耐药细胞系的 HMG-CoA 还原酶表达明显增加。此外,在氟伐他汀耐药性复制子细胞中,流出转运蛋白 P-糖蛋白的表达增加(通过 qRT-PCR 和流式细胞术确定)。这种增加的表达也导致了 calcein AM 的 P-糖蛋白介导的外排的功能转运活性增加。总之,我们证明了 HCV 复制子含有肝癌细胞中的他汀类药物耐药性是由细胞环境的变化引起的。