Fig. 7: The MiR-101–UBE2D1 axis enhances HCC chemosensitivity by increasing DNA damage.

A Western blotting analysis of UBE2D1 expression changes in SNU-739 and HCC-LM3 cells following stable transfection of miR-101 and rescue expression of UBE2D1 on this basis. B The miR-101 and UBE2D1 transcript levels were quantified by qRT-PCR in miR-101 overexpression cells and rescued cells. C, D Cell proliferation of HCC cells was analyzed in the rescue experiments following treatment of 0.5 μM cDDP or 0.5 μM 5Fu for 0, 24, 48 and 72 h respectively by CCK8 assay. E, F The cell proliferation of HCC cells was analyzed in the rescue experiments after treatment with 0.5 μM cDDP or 0.5 μM 5Fu using plate clone formation assay. G Cleaved caspase 3 expression levels were analyzed using western blotting in the rescue experiments performed on SNU-739 and HCC-LM3 cells, with or without administration of cDDP (50 μM, 24 h) or 5Fu (50 μM, 48 h). H Cell apoptosis rate was detected by flow cytometry in the rescue experiments with the cDDP or 5Fu treatment (5 μM, 24 h). I, J Detection of γ-H2AX DNA damage foci in the rescue experiments of SNU-739 and HCC-LM3 cells by western blotting and immunofluorescent staining (scale bar = 10 μm). The statistical scatter points represent the quantification of γ-H2AX foci per cell. K Comet assay was performed on the rescue experiments with or without administration of cDDP or 5Fu (10 μM, 12 h) (scale bar = 150 μm). All experiments were performed three times. Statistical significance was determined by Student’s t-test. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05.