Study On Evodiamine Arrest Cell Cycle And Induction Apoptosis Of Small Cell Lung Cancer

IntrodutionLung cancer is the most common form of cancer, accounting for 12.5% of the all new cancer cases each year around the globe. In addition, lung cancer is of the highest mortality in all cancer types. Small cell lung cancer(SCLC) is approximately 10-15% of lung cancer. Chemotherapy is the primary treatment of SCLC. However, these drugs have only limited therapeutic effect, and can cause serious side effects. Therefore, anticancer drugs with effective and less side effects are urgently need. Evodiamine(EVO) has shown cytotoxic effects in different types of human cancer cells. EVO may be a potential drug for the treatment of SCLC. But so far there is not any report on EVO for SCLC. In this study, after SCLC NCI-H446 and NCI-H1688 cells were intervented by EVO, apoptosis and cell cycle were dectected by flow cytometry, apoptosis-related proteins and genes were anlyzed by Western blot and RT-PCR, and the signal pathways of EVO arrest cell cycle and induction apoptosis of SCLC were seeked.MethodsNCI-H446 and NCI-H1688 cells were respectively divided into experimental and control groups. After cells in experimental groups were intervented by EVO, MTT method was used to detect cell survival, flow cytometry to detect cell cycle, apoptosis, intracellular reactive oxygen species(ROS), calcium and mitochondrial membrane potential changes. Apoptosis-related proteins Caspase-3, Caspase-8, Caspase-9 activities were detected by immunofluorescence analysis. Cytochrome C, Caspase-3, Caspase-8, Caspase-9, Caspase-12, FAS, Trail protein and m RNA expression were anlysized by Western blot and RT-PCR. The signal pathways of EVO arrest cell cycle and induction apoptosis of SCLC were analyzed.ResultEVO can significantly inhibit the viability of SCLC cells H446 and H1688 cell depending on the dose and time. H446 and H1688 cell cycle were arrested in G2 / M phase by EVO(P<0.05). ROS and calcium levels of cells in the experimental group were significantly increased(P<0.05), the inner mitochondrial membrane potential was significantly decreased(P<0.05). Intracellular Caspase-3, Caspase-8, Caspase-9 activities were significantly increased(P<0.05). Cytochrome C, Caspase-12, Caspase-9, Caspase-3 protein and m RNA expression levels were significantly increased(P<0.05), however, Caspase-8, FAS and Trail levels did not change significantly(P>0.05). BAX mRNA expression in the experimental group increased significantly(P<0.05), whereas the expression of BCL-2 decreased significantly(P<0.05).Conclusion(1) EVO can inhibite cell growth of SCLC H446 and H1688 cells by arrest the cell cycle in G2 / M phase.(2) EVO can inhibite cell growth of SCLC H446 and H1688 cells by inducing apoptosis.(3) EVO may induce SCLC H446 and H1688 cell apoptosis through the mitochondrial-dependent apoptotic pathway and endoplasmic reticulum pathway, rather than through the death receptor pathway.