Casticin Selective Enhance TRAIL Induced H446 Stem Cell Apoptosis

Background: Lung cancer as one of the most common malignancy worldwide. Especially in China, both male and female cancer deaths are the first one lung. Currently treatment of lung cancer is still very unsatisfactory, the 5-year survival rate of less than 15%. Numerous studies show that cancer stem cells(cancer stem cells, CSC), which accounted for some of the cells in the tumor cells, though rarely, but it is the key to tumorigenesis and development. Also contributed to the formation of tumors and tumor continuous growth, invasion, the root cause of local recurrence and distant metastasis of lung cancer have also contributed to the root cause of the ideal prognosis. And there is still no targeted eradication of drugs and treatments CSCs.Purpose: This study aims to observe the Casticin can selectively enhanced the expression of TRAIL to induce lung cancer stem cell like cells apoptosis, and to explore its mechanism of action, and provide a theoretical basis for clinical application of Casticin more fully.Methods: stem cell conditioned media culture and proliferation of stem cells in the low adhesion H446 6-well plates; TRAIL in combination with the H446 cells were cultured stem cells and ordinary divided into nine groups with different concentrations Casticin and TRAIL protein concentration and the proportion of fixed Casticin,respectively Casticin or TRAIL to act on these cells; group 1 unsortedordinary H446 cells, the medium does not add Casticin or TRAIL;Group 2 through stem cell conditioned medium of H446 sorting stem cells in culture without Casticin or TRAIL; group 3 H446 stem cells,each 2mL medium added 1μmoL of Casticin; group 4 H446 stem cells,each 2mL medium plus 3μmoL of Casticin; Group 5 for H446 stem cells,each 2mL culture base added 10μmoL of Casticin; Group 6 for H446 stem cells, each 2mL medium added 1ng of TRAIL; Group 7 for H446 stem cells, each 2mL medium added 10 ng of TRAIL; Group 8 for H446 stem cells, each 2mL medium was added 100 ng of TRAIL; 9th group H446 stem cells, the culture medium was added per 2mL of TRAIL and3 umo L Casticin 10 ng of; were cultured for 24 hours, and then compare the size and number of balls into each group in a microscope, and by ELISA, FACS, Western-bolt and other experimental methods to detect apoptosis of H446 stem cells. With lentiviral overexpression and knockdown FoxM1 way to explore Casticin selectively enhances TRAIL-induced apoptosis in H446 stem cells.Results: The use of stem cell conditioned medium obtained by culturing stem cells H446 by protein immunoblot(Western-bolt), flow cytometry can be used for follow-up study, the stem cell marker UPAR H446 cells were significantly higher than normal; and with different concentrations Casticin TRAIL protein alone and combined effects on H446 stem cells, the results show, Casticin with TRAIL has the ability to number H446 stem cells into a ball and a ball the size of the inhibition and Casticin can selectively enhance the role of TRAIL, both of which can be combined to more significant inhibition H446 stem cells balling capacity; found Casticin enhanced TRAIL selectively induces apoptosis H446 stem cells, the anti-apoptotic protein c-Filp, such as decreased expression of P56 by Western-bolt, ELISA, FACS and otherexperimental methods; and using lentivirus overexpression and knockdown way to determine FoxM1 is Casticin selective enhancement of stem cells TRAIL-induced apoptosis in H446 important genes.Conclusion: The first time that both promote H446 Casticin stem cell apoptosis; and confirmed Casticin TRAIL caused by selectively enhancing H446 stem cell apoptosis; by lentiviral transfection technique proved Casticin is selectively enhances TRAIL-induced H446 apoptosis through FoxM1 genes.