Ellagic On The Biological Effects Of Myeloma SP2/0Cells

OBJECTIVE: The research with mouse myeloma SP2/0cells as research subjects. Use of in vitro cell culture and molecular biology techniques,and other research methods, Explore gallnut extract ellagic acid on multiplemyeloma cells proliferation, apoptosis and the impact of cell cycle, And detect the expression of tumor associated molecules-COX-2which Closely related to cancer occurrence and development. Through the analysis of ellagicacid on mouse myeloma SP2/0cell biology and molecular mechanisms ofaction to provide an experimental basis for clinical surgery MBD, effectiveremoval of the lesion myeloma cells.Methods: Mouse myeloma SP2/0cells cultured in vitro. Establish three drug group and a control group andLadder-like drug concentration, Ellagic acid control group without intervention,Three experimental groups were20，40and60μg/ml ellagic acid intervention.1、using an inverted microscope, different concentrations of ellagic acid treatment in the experimental group and the control group myeloma SP2 /0cells48h after changes in cell morphology and photographed.2、with20，40and60μg/ml ellagic acid treatment of myeloma SP2/0cells48h, using Hoechst33258staining method, and then inverted fluorescence microscope contrast apoptosis in the experimental group and the control group of cells and photographed.3、Using the above concentrations of ellagic acid treatment after48h, through Cell proliferation assay detects SP2/0cell proliferation and its viability.4、Collecting SP2/0cells which have been treated by above four concentrations groups of ellagic acid after24h. then by flowcytometry to determine the effect of ellagic acid on myeloma SP2/0earlyapoptotic cells.5、The above concentration drug has treated SP2/0cellsfor48h，then，determine for each cell rate of the cell cycle by flow cytometry.6、By Western blotting technique to detect the expression of Tumor appreciation and apoptosis-related proteins---COX-2changes in SP2/0cellswhich have been treated by different concentrations of drugs.Results:1、Different concentrations of the drug treatment after48h SP2/0cells compared with control group，with increasing concentrations of ellagic acid，decreasing the number of adherent and active cells,meanwhile broth floating dead cells growing.2、Hoechst33258stained cells were observed under an inverted fluorescence microscope，Nuclear group showed diffuse, uniform blue fluorescence, with the increase of drug concentration in the experimental groupthe number of stained cells in the cell decreases and the dense stain visiblefluorescence. The cell proliferation assay found that20，40and60μg/ml were incubated ellagic acid group cell growth inhibition rate was (21.18±5.92)%,(44.58±3.43)%and (70.15±2.90)%respectively,there was significant difference compared with the control group (P <0.01).4、The rate of detection of early apoptotic cells, experimental group SP2/0cell apoptosis rate were (9.60±0.56)%,(19.30±1.51)%and (35.10±5.26)%, comparedwith the control group (3.23±0.85)%, p <0.01, the difference statisticallysignificant.5、Cell cycle analysis showed that ellagic acid incubation myeloma SP2/0cell lines after48h, cell cycle arrest in G1phase, G1phase cell percentage were (55.21.±3.01)%,(64.48±0.43)%,(75.10±2.46)%, and compared with the control group (34.04±1.74)%, the difference was statistically significant (P <0.01).6、Western blotting technique detected,protein expression of COX-2decreased consistently with increasing drug concentration and showed a significant concentration-dependent manner.Conclusions: Gallnut extract ellagic acid can inhibit mouse myeloma SP2/0cell proliferation, promote apoptosis may be blocked in G1phase. The mechanism of this process may relate to the expression of COX-2down in the SP2/0cells. Description ellagic acid has potential anti-myeloma biological functions.