Experimental Study On Reactive Oxygen Species Mediated Apoptosis Of Human Colon Cancer HT-29Cells Induced By5,7-dimethoxyflavone

ObjectiveTo investigate whether5,7-dimethoxyflavone (5,7-DMF) induces apoptosis ofhuman colon cancer HT-29cells and explore whether this mechanism is involved ininduction of reactive oxygen species (ROS) generation.MethodsHuman colon cancer HT-29cell line was cultured in vitro. The histone/DNAfragment in HT-29cells was evaluated using enzyme linked immunosorbent assay(ELISA). The cell apoptotic rate of HT-29cells was analyzed by flow cytometry(FCM) using propidium iodide (PI) staining. The generation of ROS was analyzed byFCM using the fluorescent probe2’7’-dichlorofluoresein diacetate (DCFH-DA).ResultsThe results by ELISA showed that5,7-DMF in a concentration-dependentlyincreased histone/DNA fragments in human colon cancer HT-29cells(P＜0.05).FCM with PI staining indicated that5,7-DMF significantly promoted the apoptoticcell death of HT-29cells, in a concentration-dependent manner(P＜0.05). FCMusing DCFH-DA found that5,7-DMF effectively induced the generation of ROS inHT-29cells. In addition, pretreatment with10mmol/L N-acetylcysteine efficientlyblocked induction of ROS production and apoptosis by5,7-DMF in HT-29cells. Conclusion1.5,7-DMF possesses induction of apoptosis in human colon cancer HT-29cell line,in a concentration-dependent manner.2.5,7-DMF effectively increased the generation of ROS of human colon cancerHT-29cells.3. The induction of apoptosis by5,7-DMF in human colon cancer HT-29cells wasassociated with induction of ROS production.