Dihydroartemisinin combined with cisplatinum induced apoptosis and related mechanism research in human lung adenocarcinoma H1299 cells

Dihydroartemisinin (DHA), a kind of artemisinin derivatives, its activity in anti-cancer was extensively studied in recent years. There is not yet related research about dihydroartemisinin puls cisplatinum (Cis) in treatment of a human lung adenocarcinoma cancer cell line (H1299 cells) so far. This experiment is designed to study the apoptotic effect induced by DHA and Cis to H1299 cells. MTT assays, nuclear staining with Hoechst 33258 and flow cytometry were used to evaluate the apoptosis status. The expression of apoptotic-associated proteins was assessed by Western blotting and RT-qPCR. DHA and Cis pro-apoptotic activity of alone and combined administration enhanced as drug concentration increased or elongation of time. Flow cytometry revealed that the apoptosis rate of the singnal medicine group and the combined medicine group were higher than the control group. The result of RT-qPCR reveal that drug combination of Ct value about caspase-3,-8,-9 are significantly higher than monotherapy. The combined treatment with two drugs significantly enhanced caspase-8,-9,-3,-6,-7 and PARP activation, indicating that the caspase-8 participated extrinsic apoptosis pathway and the caspase-9 participated intrinsic apoptosis pathway played improtant roles respectively in the synergistic effect. In general, this study reveals the obvious synergistic action of the combined treatment with DHA and Cis in inducing apoptosis of H1299 cells by the extrinsic and intrinsic apoptosis pathway.