Study Of Effect Of Triptolide On Proliferation And Apoptosis On U2OS Cells

Osteosarcoma is one of the most common primary malignant bone tumors,the bulk of people who get osteogenesis sarcoma are below age 21 and it mainly happened in the metaphysis of the bone.In addition to the individual study,clinical research is still in the blanking period in the molecular targeted drugs of osteosarcoma.So the cure rate of osteosarcoma is low and has a big harm to adolescent health.So efforts should be made to develop new treatment strategies and develop new drugs,in which the molecular targeted therapy may become important research direction.With the development of modern molecular biology techniques,we have a deeper understanding in cell proliferation apoptosis,tumor development and treatment,which provide us the scientific and objective of modern means and methods,also provides some new train of thoughts in the research of antineoplastic mechanism of traditional Chinese medicine.In our study,we adopted the small molecular triptolide to treat U2OS cells,and then observed the effects of medicine on growth,proliferation,apoptosis of U2OS cells,and detected the change of Caspase8/9 and PARP level.We proved that the medicine can inhibit the growth and proliferation of U2OS cells,that provided the cytological basis for the follow-up drug anti-cancer research.In our study,we adopted 5nmol/L、10nmol/L and 25nmol/L triptolide to treat U2OS cells,then used CCK8 colorimeter and trypan blue staining,counted cell numbers and drew growth curve to detect the effect of medicine on growth of U2OS cells.We double dyed cells and detected the apoptosis by fluorescence microscope and flow cytometer.P53 level and caspase signal transduction were measured by western blot.The result of growth curve showed that 5nM triptolide can inhibit growth of U2OS cells in a certain extent,and 25nM triptolide completely inhibited growth of U2OS cells.IC50 result detecting by CCK8 colorimeter showed that IC50 of U2OS cells treated by triptolide is 18.4nM.Treatrd with triptolide after 48h,the cells apoptosis happened,and with the increase of drug concebtration,the number of apoptosis is also increased.Still treatrd with triptolide after 48h,analyzed the testing result of the cell cycle,can know that triptolide can make the cell growth stop in G1 phase.By colony formation assay,we find that triptolide has obvious inhibition for the growth of cell proliferation.Triptolide has significant regulation to the related proteins of the caspase signal transduction.Therefore,we arrival at a conclusion,triptolide can inhibit growth,proliferation of U2OS cells,induce apoptosis and cell cycle arrest happened in G1 phase,and triptolide can regulate the expression of apoptosis signaling pathways related proteins.