The Research Of Matrine On The Suppression And Its Mechanism Of Raji Cells

Objective:Burkitt lymphoma is a malignant tumor of hematopoietic system. Raji cell is a common Burkitt lymphoma cell used in basic research. Chemotherapy is a prevalent method for Burkitt lymphoma but the drugs are selectively limited and have toxic side effect which will cause damage to immune and hemopoietic system. Therefore, it is of importance to find drugs with little side effects and high selectivity in order to improve of the clinical efficacy. Matrine, a alkaloid separated from leguminous plant, has an anti-tumor activity mainly by suppressing cell proliferation according to some research reports. But no research reported to the effect of matrine on Raji cells.In this study, we observed the impact of Mat on the morphology and cell cycle of Raji cells to explore whether Mat could suppress the proliferation of Raji cells. We observed mRNA and protein expreesion of cyclinE and its inhibitor p27kip1 in Raji cells, to explore possible mechanisms of Raji proliferation.Methods: 1. Inverted microscope was used to observe the changes in cell morphology.2. The proliferation inhibition rates of Raji cells were examined by MTT assay.3. Flow cytometry was used to observe the effect of cell cycle distribution of Raji cells.4. The expression and distribution change of the cyclinE,p27kipl,proteins were detected by immunohistochemical staining.5. cyclinE mRNA and p27kipl mRNA expression of Raji cells were detected by RT–PCR.6.The changes of cyclinE,p27kipl proteins were detected by Western blot.Results: 1. Inverted microscope observation of Raji cells, after being treated with 0.4 mg/ml,0.8 mg/ml,1.2 mg/ml,1.6 mg/ml,2.0 mg/ml matrine for 48 hours, showed reduction of volume and karyoplasmic ratio, compared with the control group.2. After the 24 hours, 48 hours and 72 hours of treatment with 0.4 mg/ml,0.8 mg/ml,1.2 mg/ml,1.6 mg/ml,2.0 mg/ml matrine, MTT assay showed that matrine can inhibit the proliferation of the Raji cell,and the inhibition was dependent on both dose and time.The IC50 of 48 hours was 0.8712 mg/ml.3. After the 48 hours of treatment with 0.4 mg/ml and 0.8 mg/ml matrine,the number of cells in G1–phase increased,whereas the number in S-phase decreased.4. After the immunocytochemical staining test, showed the cyclinE and p27kipl were located in the cell nucleus and plasma. After the 48 hours of treatment with 0.4 mg/ml and 0.8 mg/ml matrine,the expression of cyclinE was down-regulated,and whereas p27kipl was up–regulated by immunocytochemical method.5. After the 48 hours of treatment with 0.4 mg/ml and 0.8 mg/ml matrine,the cyclinE mRNA expression in Raji cells was decreased and p27kipl mRNA expression in Raji cells was increase, showed by RT-PCR.6. After the 48 hours of treatment with 0.4 mg/ml and 0.8 mg/ml matrine,the protein expression of cyclinE was down-regulated,whereas p27kipl was up–regulated by westem blot.Conclusions: 1. Matrine can inhibit the proliferation of the Raji cells,and the inhibition was dependent on both dose and time.2. Matrine can affect Raji cells cycle distribution,inhibiting cell proliferation.3. The mechanism of inducing Raji ceils differentiation by matrine might be related to the mRNA and protein expression of cyclinE was down-regulated,whereas p27kipl was up–regulated.