Effects of ASX on HCC cell proliferation. (A) LM3 and SMMC-7721 cells were treated with DMSO and ASX (100 μM, 200 μM, 300 μM) for 72 h. The effect of different concentrations of ASX on growth was lower than of DMSO and was dose-dependent; (B) LM3 and SMMC-7721 cells were treated with ASX (50−300 μM) and evaluated at 24 h, 48 h, and 72 h. The data showed that ASX reduced HCC cell proliferation in a time-dependent manner; (C) The mRNA levels of PCNA were determined by RT-PCR (n = 3, *,#,+
p < 0.05 for ASX versus DMSO); (D) The protein levels of PCNA were assessed by western blotting and the relative band intensities of PCNA were calculated using the Odyssey two-color infrared laser imaging system (n = 3, *,#,+
p < 0.05 for ASX versus DMSO).