| The traditional Chinese medicine plays an important role inthe treatment of malignant tumors. The experiment is to studythe anti-tumor effects exerted by honeycomb in vitro.Material and Methods: The liver tumor cell SMMC-7721were cultured in RPMI-1640 media with 10% FCS, 100u/mlpenicillin and streptomycin on the condition of 37℃ 5% CO2.There were two group in the experiment. One is experimentgroup which the different concentration of honeycomb waterextracts was dissolved in the culture media; the other is controlgroup which the same volume of aseptic distilled water as thatof experiment group was dissolved in culture media. 1. MTTassay: The suspended cells obtained from the log phase culturedcells were incubated into 96-well plates. 24 hours later, the drugwas added into media at the concentration of1:40,1:80,1:160,1:320. At the end of 48 hours and 72 hours,MTT was added, then to test. 2. Growth curves: Single cell.suspension 1×104/ml was incubated into 24 well plates, thewater extracts of honeycomb were added at the concentration of1:80, then to draw the cell growth curves. 3. Colony-formingtest: Single cell suspension 500 cells/ml was incubated into 6well plates, added 20ul drug in the experiment group and 20ulwater in the control group, cultured for 8 days, then count thenumber of colony. 4. Cell cycle analysis: The cell suspensionswere cultured in 50 cm2 glass flasks, 5 ml each flask, 20ul drugand 20ul water were added in experiment and control grouprespectively After 24 hours and 48 hours medication, performedDNA staining then examined them with flow cytometer, andanalyzed the result by Modifit software. 5. Apoptosis detection:l X l0e cells were planted into the flask. After adding 20ul drugand 20ul water, the cells were cultured fOr 24 and 48 hours, thendetection was carried with the flow cytometer according to theAnnexin V/Pi apoptosis kit direction, and the resuIts wereanalyzed by CellQuest software.Result8: l. MTT showed the honeycomb water extractssignificantly suppressed the growth of liver tumor cells, and thedose-effect dependence, time-effect dependence relationshipwere existed. At 48 hours after medication, the cell growthinhibition rate was 85.4% in the l:40 diluted medicine group,P<0.01; At 72 hours the rate was 97.3%, P<0.0l; 2.Growthcurve showed the cells of experiment group grew slowly; 3.CoIony forming test indicated that the cell colony fOrming wassignificantly inhibited by the honeycomb. The forming rate wasl8.6% f2.42% in the experiment group, 58.6%t l5.0% in thecontrol group, P=0.0l2; 4.CelI cyc1e anaIysis showed: In theexperiment group, at 48 hours, G0-Gl phase celI was 85.7%which higher than that in the control group, P<0.0l, S phasecells were l l .6l% and G2-M phase cells were 2.68%, lower thanthese in the control group, P<0.0l; 5. Apoptosis detectionresults suggested the honeycomb induced the apoptosis of Iivertumor cells. 48 hours after treatment, the cell apoptosis rate wasl0.5l% i 0.53% in the experiment group, and 2.09%f0.79% inthe control group, P<0.05.Conclusion:The honeycomb water extracts influenced the S-phase ofliver tumor cell SMMC-7721, blocked the cells going into G2-M phase, induced the apoptosis of the tumor cells and inhibitedthe cuItured tumor cells proliferation in vitro. |
PDF Full Text Request