Influences of withaferin A on proliferation, apoptosis and PI3K/Akt signaling pathway of non-small cell lung cancer A549 cell

CAI Yong; WANG Jiying

Abstract：Objective To explore the influences of withaferin A ( WFA) on proliferation, apoptosis and PI3K/Akt signaling pathway of non-small cell lung cancer (NSCLC) A549 cell. Methods The A549 cells were treated with different concentrations of WFA ( 0, 2.5, 5.0, 10.0, 20.0 μmol/L) . The MTT assay was used to measure the proliferation inhibition rates at 24, 48, 72 and 96h treated with different concentrations of WFA. The Hoechst staining and Annexin V-FITC/PI double staining were employed to de-tect cell apoptosis after treatment with different concentrations of WFA. The cycle distribution at 48h after treatment with WFA was de-tected by flow cytometry. The Western blotting was used to measure the protein levels of apoptosis-related genes ( Bcl-2, Bax and Cleaved caspase-3) , Akt and its phosphorylated form ( P-Akt) . Results WFA could increase the proliferation inhibition rates in a dose-and time-dependent manner( P<0.05) . The apoptotic indices of A549 cells after treatment with different concentrations of WFA (0, 2.5, 5.0, 10.0, 20.0 μmol/L) were 2.75±0.64, 4.6±1.36, 9.75±2.78, 12.92±3.42 and 18.68±4.31 with significant differences ( P<0.05) . In addition to 2.5μmol/L group, the early and late apoptosis rates, protein levels of apoptosis-promoting genes ( Bax and Cleaved caspase-3) and the cell proportion in G0/G1 phase of the remaining concentration groups were higher than those of 0μmo/L group(P<0.05).The p-Akt/Akt value decreased with increasing concentration and the differences were statistically significant among concentrations ( P<0.05) . Conclusion WFA can inhibit the proliferation and apoptosis of A549 cell possibly by inhibiting the activation of PI3K/Akt pathway activation.