Fig. 3.

ABCC5 mediates the acquired resistance to sorafenib in vitro. (A, B) qRT-PCR and Western blot analysis detected the expression of ABCC5 in HCC cells after knockdown and overexpression of ABCC5. (C) HuH7 cells were transfected with LV-ABCC5 and were treated with 3 μM sorafenib for 24 h. UPLC8-MS/MS was used to examine the intercellular concentration of sorafenib. The formation rates of M3, 4, 5 were expressed as relative ratios (Mean ± SD). (D) HuH7-SR cells were treated with 3 μM sorafenib for 24 h, and the intercellular concentration of sorafenib was determined by UPLC8-MS/MS. The formation rates of M3, 4, 5 were expressed as relative ratios (Mean ± SD). (E) The effect of the ABCC inhibitor MK571 (10 μM) on sorafenib resistance was evaluated in HuH7 and Sk-Hep-1 cells by CCK-8 assays. (F) qRT-PCR and Western blot analysis detected the expression of ABCC5 in HuH7 cells and ABCC5-knockdown HuH7-SR cells. (G) Clonogenic cell survival assay. HuH7-SR cells were treated with sorafenib (10 µM) for 24 h, then 1000 cells were plated into 6-well plates. Colonies were visualized by crystal violet staining two weeks later. # P > 0.05, * P < 0.05, ** P < 0.01.