Study On The Effect Of Neferine On Anticancer Effect Of Pirarubicin In Vitro

Objective:We choose Pheiffer cells of diffuse large B-cell lymphoma as target cell.To observe the effect of neferine on anticancer effect of pirarubicin in vitro. With the result of this study,we can research the effect of chemosensitizer on primary orbital lymphoma in the future.Methods:1. The DLBCL cells are reanimated, observe the proliferative status and morphological characteristic.2. DLBCL cells were plated in 96-well plates and cultured in medium with 10% fetal bovine serum for 24h. Then DLBCL cells were divided into control group and test groups of Nef at different density(0,5, 10μmol/ml) at random. After the cells were cultured for 48h respectively, the morphological change was observed, cell proliferation is determined by MTT3. DLBCL cells were plated in 96-well plates and cultured in medium with 10% fetal bovine serum for 24h. Then DLBCL cells were divided into control group and test groups of THP at different density(0.0275,0.055,0.11,0.22,0.44μg/ml) with Nef at different density(0,5,10μmol/ml)at random. After the cells were cultured for 48h respectively, the morphological change was observed, cell proliferation is determined by MTT.4. Analyze the effect of different density of THP and/or Nef on the cell cycle and apoptosisi of DLBCL cells by FCM.Results:1. DLBCL cells developed well and passed stably. The shape of cell is round or ellipse.2. Nef at 5, 10μmol/ml has no obvious cell toxicity, the inhibition rate were all less than 15%.3. After cultured in the medium supplemented with different density of THP for 48h, the proliferation of the cells was inhibited at different degree(P<0.05). Nef at 5, 10μmol/ml significantly decreased the IC50 THP to DLBCL cells.4. The result of FCM indicates that the cell eycle of DLBCL cells treated by THP has changed. The proportion of cells in stage G1 increased, conversely, the proportion of cells in stage S and G2 decreased(P<0.05).The THP induced apoptosis in DLBCL cells was enhanced by 10μmol/ml Nef.Conclusion:1. Nef at 5,10μmol/ml that has no obvious cell toxicity can significantly decreased the IC50 THP to DLBCL cells.2. THP intervene the cell cycle of DLBCL cells, the proportion of cells in stage G1 increased, as a result, the proliferation of cell is inhibited. The THP induced apoptosis in DLBCL cells was enhanced by 10μmol/ml Nef.3. Nef is able to potentiate the anticancer effect of THP and may be a chemosensitizer.