Figure 7. Aurora-A inhibits chemotherapy-induced apoptosis in HCC cells by upregulation of miR-21.

(A) Western blotting detection of apoptosis-related proteins (PTEN, p-Akt, total Akt, Bcl-2, Bax, cleaved caspase-3 and total caspase-3) in SMMC-7721/shAurora-A (or SMMC-7721/shcontrol) or HepG2/Aurora-A (or HepG2/control) cells. (B) Western blotting detection of total Aurora-A, p-Aurora-A and above apoptosis-related proteins in SMMC-7721 cells treated with various concentrations of MLN8237 (0.0, 0.5, 1.0 and 2.0 nM) for 24h or lengths (0, 6 and 12h) of MLN8237 (1.5 nM). (C) Flow cytometry detection of apoptosis in SMMC-7721 cells treated with various concentrations of MLN8237 (0.0, 0.5, 1.0 and 2.0 nM) for 24h or lengths (0, 6 and 12h) of MLN8237 (1.5 nM). (D) 24h after HCC cells (HepG2 and SMMC-7721) were treated with PBS, ADR (0.5 μg/ml) or CDDP (1.0 μg/ml), qRT-PCR detection of miR-21 expression. (E) qRT-PCR detection of miR-21 expression in SMMC-7721/shAurora-A (or SMMC-7721/shcontrol) or HepG2/Aurora-A (HepG2/control) cells, respectively. (F) qRT-PCR detection of miR-21 expression in SMMC-7721 cells treated with various concentrations of MLN8237 (0.0, 0.5, 1.0 and 2.0 nM) for 24h or lengths (0, 6 and 12h) of MLN8237 (1.5 nM). (G) MTT analysis of IC₅₀ values of ADR or CDDP in SMMC-7721/shAurora-A or SMMC-7721/shcontrol cells or co-transfection with miR-21 or miR-NC mimics, or in HepG2/Aurora-A or HepG2/control or or co-transfection with miR-21 or miR-NC inhibitor, respectively. (H) Western blotting detection of above apoptosis-related proteins in SMMC-7721/shAurora-A cells or co-transfection with miR-21 or miR-NC mimics. (I) Western blotting detection of above apoptosis-related proteins in HepG2/Aurora-A cells or co-transfection with miR-21 or miR-NC inhibitor. GAPDH or U6 was used as an internal control. Data were presented as mean ± SD of at least three independent experiments. N.S, P>0.05; *P<0.05; **P<0.01.