CA derivatives significantly inhibited the proliferation of human CRC SW-480 cells <i>in vitro</i>.

<p>(A) Human CRC SW-480 cells were cultured in RPMI-1640 medium with CAPE and CAPPE (at concentrations of 0, 5, 10, 20, 50 and 100 µM) in the presence or absence of compound C (10 µM) for 24 h. Transfections of constitutively active Akt (Myr-Akt1) and empty vector (pcDNA3) were conducted before the treatment of CA derivatives. The cell proliferation was measured by MTT assay as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099631#s2" target="_blank">Materials and Methods</a>. Data are the mean ± SD (standard deviation) of three independent experiments. The different symbols (??? for CAPE and ▵ for CAPPE) represent a statistically significant difference compared to the CA derivative -untreated control group in each group, respectively, at P<0.05. The different symbols (# for CAPE_Akt, § for CAPE_compound C, ▴ for CAPPE_Akt, and ▪ for CAPPE_compound C) represent a statistically significant difference compared to each corresponding CA derivative- treated control group in each dosage subgroup, respectively, at P<0.05. (B–C) Cytoplasmic proteins were prepared for Western blotting analysis using monoclonal antibodies against anti-phosphorylation Akt (S473), total-Akt, anti-phosphorylation AMPKα (T172) and total-AMPKα.