The Study Of Berbamine Inducing On Lymphoma Raji Cell's Apoptosis

Non-Hodgkin's lymphoma(NHL) is one of malignant tumors sourced from lymph nodes and(or) extra-nodal lymphoid tissue,NHL Patients usually showed good response on traditional chemotherapy(such as COP,CHOP,etc.),but most of them often easily recurred after relief for a few months even several years.The patients could achieve long- term relief time after stronger chemotherapy,but the stronger chemotherapy drugs followed the more toxicity and higher mortality rate,which made new therapy for the NHL become a hot spot in recent years.The berbamine has shown obvious anti-tumor effect on a variety of malignant tumors.The mechanism are related to change the molecular conformation of calmodulin, and inhibit the target enzyme which are regulated by calmodulin,so as to affect the metabolism, cycle growth and other biological activity of tumor cell.It has tolerable side effect and no cross-resistant drug with other chemodrugs.To provide theoretical proof for new combination chemotherapy programs in clinic, we treated human lymphoma cells Raji with berbamine,and observed the changes of the cells proliferation and cells apoptosis rate.Wright-Giemsa dyeing assay was used to observe apoptosis morphology of lymphoma cells.Detecting proliferation of the cells with berbamine by the method of MTS.Flow cytometry(FCM) and DNA ladder was used to detect apoptosis of lymphoma cells.western blot was used for detecting PARP(poly ADP-ribose polymerase).The result shows that the cell viabilites were significantly inhibited by BBM at the concentration of 0-32μg/ml,The cell survival rate was decreased with the increasing concentration of BBM.Furthermore,BBM inhibited cell growth in a time dependent manner. If treated at the same concentration of BBM,fewer cells would survive when the cells were treated for an increasing time.The 24h,48h,72h IC50 of BBM on Raji cell were 16.01μg/ml,11.74μg/ml,9.90μg/ml.After treated by BBM for 24h,Raji cells observed by Wright-Giemsa stain,showed a typical apoptotic morphology,nuclear fragmentation and apoptotic body.Typical DNA ladders were also seen in Raji cells treated by BBM for 24h.With the concentration of BBM increasing,The DNA ladder became more significant.BBM could induce apoptosis in Raji cells,as measured by AV-PI assay. After treated by 1,2,4,8,16,24,32μg/ml BBM for 24h,the Raji cells in early apoptosis were38.89±2.11%,41.43±3.45%,44.54±3.02%, 47.09±2.69%,54.20±5.67%,57.47±6.99%,60.87±5.38%,Furthermore,PAR P were cleaved and activated,as measured by western blot in Raji cells treated by BBM。In conelusion,berbamine can inhibit Raji cells proliferation,induce cells apoptosis,which may through caspase way.