The Inhibitory Effect Of Glaucocalyxin A On Human Lung Cancer A549Cell And Its Mechanism

Objective： To investigate the antiprolifrative effect and the mechanism ofGlaucocalyxin A on human lung cancer A549cells.Methods: Human lung carcinoma A549cell was treated with differentconcentrations of GLA（0,5,10,20μmol/L）, investigate the cytoxicity of GLA by MTTassay. Morphological and biochemical changes of apoptosis were investigated usingHoechst33258staining and TEM. The ratio of apoptotic cells was measured by flowcytometry using AnnexinV/PI staining.The change of cell cycle was also measured byflow cytometry using PI staining.25mg/kg).Western bloting was used to detect theprotein level of caspase-3、caspase-8and Parp. RT-PCR to detect the gene changes ofcaspase-3、caspase-8and Parp. Real time PCR to detect the level of microRNA30A.Meanwhile,flow cytometry was used to detect the effect of3-MA on apoptosisinduced by GLA.Results: Compared with control group, GLA displayed potent cytotoxic activityagainst human lung cancer A549cell. GLA induced apoptosis in A549cells, and the ratioof apoptosis cells increased (P<0.01). Apoptosis morphological changes were observedon A549cells after GLA treatment by Hoechst33258fluorochrome staining、TEM andSEM.Sub-G1phase peak was appeared and cell cycle was arrested in S phase afterGlaucocalyxin A treatment. The expression levels of caspase-3、caspase-8and ParpmRNA were downregulated(P<0.05, P<0.01). Real Time PCR detection miRNA-30aexpression was significantly lower (P<0.05, P<0.01); Western blot detection ofcaspase-3、caspase-8and Parp were upregulated(P<0.05, P<0.01).Conclusion: GLA can inhibit A549cell proliferation and induce cell apoptosis.GLAalso can induce cell cycle arrest. GLA induced apoptosis in human lung cancer A549 cells trough activating caspase8patnway.3-MA could increase the ability of GLA inhibitA549cell.