Antiproliferative Effect of Ligustilide on Human Lung Adenocarcinoma A549 Cells / 中国中医药信息杂志

ABSTRACT

Objective To investigate the effects of ligustilide on the proliferation of human lung adenocarcinoma A549 cells and its mechanism. Methods CCK-8 method was used to detect the effects of ligustilide on activity of A549 cells. Apoptosis rate was measured by TUNEL. Nuclear morphological changes of A549 cells were observed by fluorescence microscope after staining by Hoechst 33258. ELISA was used to detect the VEGF levels after incubation with ligustilide. Western blot was used to detect the expression of NF-κB, Bcl-2, Bax and the protein expression of phosphorylation of JNK. Results A549 cell activity was significantly inhibited after incubated with 15, 30, 60, 120, 180μmol/L ligustilide for 12, 24, 48 h (P<0.05, P<0.01, P<0.001) in a dose and time-dependent manner (P<0.05, P<0.01);Apoptosis rate increased by 30, 60, 120 μmol/L ligustilide for 12, 24, 48 h in a dose and time-dependent manner;A549 cells treated with ligustilide for 48 h appeared the apoptosis forms, including nuclear shrinkage, beating, strong blue fluorescence staining, and the chromatin divided producing apoptotic body;30, 60, 120μmol/L ligustilide increased the expression of p65 subunit of NF-κB protein in the nuclei and the phosphorylation levels of JNK protein, reduce the expression of suppression apoptosis protein Bcl-2, and raised the expression of pro-apoptotic protein Bax (P<0.05, P<0.01, P<0.001). Conclusion By inhibiting the expression of VEGF, ligustilide can inhibit the formation of tumor angiogenesis. By regulating the expression of NF-κB and the phosphorylation levels of JNK protein and reducing the ratio of Bcl-2/Bax, ligustilide can promote tumor cell apoptosis.