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Multidrug resistance protein 4 is a critical protein associated with the antiviral efficacy of nucleos(t)ide analogues
Author(s) -
Liu Wei,
Song Hongxuan,
Chen Qian,
Xu Chunli,
Zhang Wenjing,
Liu Yutian,
Wang Baoju,
Xu Dongping,
Lu Mengji,
Yang Dongliang,
Zheng Xin
Publication year - 2016
Publication title -
liver international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.873
H-Index - 110
eISSN - 1478-3231
pISSN - 1478-3223
DOI - 10.1111/liv.13104
Subject(s) - multiple drug resistance , peripheral blood mononuclear cell , hepatitis b virus , microbiology and biotechnology , intracellular , real time polymerase chain reaction , biology , blot , extracellular , cell culture , drug resistance , virology , virus , in vitro , gene , biochemistry , genetics
Background & Aims Multidrug resistance protein 4 ( MRP 4) has been associated with nucleos(t)ide analogue ( NA ) antiretroviral therapy failure, though is unclear if MRP 4 is also correlated with the failure of anti‐hepatitis B virus ( HBV ) therapy. Methods Multidrug resistance protein 4 expression in human peripheral blood mononuclear cells ( PBMC s), liver tissues and human hepatoma cell lines was detected by real‐time polymerase chain reaction ( PCR ), western blotting and immunohistochemistry assays. Supernatant and intracellular HBV DNA levels of MRP 4‐overexpressing or silenced HepG2.4D14 (wild‐type) and HepG2.A64 (entecavir‐resistant mutant) cells were measured by quantitative PCR . NA concentrations and HBV mutational analysis were assessed by liquid chromatography/mass spectrometry assays and DNA sequencing. Multivariate analysis was used to assess predictive factors for treatment failure. Results High expression of MRP 4 was found in hepatoma cell lines and PBMC s, and up‐ or down‐regulation of MRP 4 expression altered the susceptibility of cells to NA s. MRP inhibitors increased NA intracellular accumulation and decreased extracellular levels. Moreover, MRP 4 expression in PBMC s was correlated with that in paired liver tissues. Furthermore, multivariate analysis showed MRP 4 mRNA expression to be an independent predictor of NA treatment failure. Conclusions Multidrug resistance protein 4 is a critical protein associated with the antiviral efficacy of NA s, and combination therapy of NA and MRP inhibitors could reduce the dosage for long‐term NA use. This is the first report to demonstrate that MRP 4 expression is an important factor predicting treatment failure in chronic hepatitis B patients and will provide a potential therapeutic target against HBV .