ABSTRACT
Objective To determine the effect of apigenin on inducing apoptosis and inhibiting proliferation of non-small cell lung cancer cell line NCI-H1650. Methods NCI-H1650 cell line was cultured routinely in vitro, with blank control group and different concentrations of apigenin (10, 20, 40, 80 μmol?L-1). The blank control group was RPMI-1640 solution without apigenin. Cell proliferation was detected by MTT. Hoechst 33258 was applied to observe morphological changes of the apoptotic cells after treatment of different concentrations of apigenin. Flow cytometry AnnexinV-FITC/PI double staining method was used to determine cell apoptosis rate. The expression levels of apoptosis-related signaling molecules Bax and Bcl-2 protein were performed by Western blotting. Results MTT showed that apigenin inhibited proliferation of NCI-H1650 cell line in a concentration-and time-dependent manner (P<0.01).Hoechst 33258 nuclear staining showed typical characteristics of apoptosis in certain concentration-dependent manner, such as nuclear condensation and appearance of apoptotic bodies. In addition, the results of flow cytometry staining indicated that the apoptotic rate of NCI-H1650 cells cultured with blank control group and different concentrations of apigenin (10, 20, 40, 80 μmol?L-1) for 48 h was (5.00±0.33)%, (18.05±4.67)%, (21.48± 1.95)%, (43.24±1.11)%, (66.23±3.65)%, respectively (P<0.01).Western blotting results showed that the expression levels of Bax increased with increasing of the drug concentration, but Bcl-2 decreased with increasing of the drug concentration. Conclusion Apigenin can inhibit the proliferation and induce apoptosis of NCI-H1650 cells in non-small cell lung cancer. Mechanisms may be related to increase of the expression of apoptosis related protein Bax and decrease of the expression of anti apoptosis protein Bcl-2.
ABSTRACT
<p><b>OBJECTIVE</b>To study the effect of apigenin on the proliferation and apoptosis of human lung cancer cell line NCI-H460.</p><p><b>METHODS</b>NCI-H460 cells were cultured with different concentrations of apigenin, and MTT assay was used to evaluate the cell inhibition rates. Apoptosis of NCI-H460 cells was observed under a fluorescence microscope with Hoechst 33258 staining and quantified by flow cytometry using annexin V-FITC/PI stain. The expressions of apoptosis-related proteins Bax, Bcl-2 and caspase-3 were analyzed by Western blotting.</p><p><b>RESULTS</b>Apigenin causes concentration- and time-dependent inhibition of the proliferation of the cells. NCI-H460 cells treated with apigenin showed significant morphological changes of apoptosis, and the cell apoptotic rates increased as apigenin concentration increased. Western blotting demonstrated that apigenin increased the protein levels of Bax and caspase-3 and reduced the protein expression of Bcl-2.</p><p><b>CONCLUSION</b>Apigenin can inhibit the proliferation and induce apoptosis of NCI-H460 cells possibly by up-regulating expression of Bax and caspase-3 and down-regulating the expression of Bcl-2.</p>