Clematis Polysaccharide Of Tongue Squamous Cell Carcinoma Cell Growth Inhibition Studies

Objective:T0 study the inhibitory effect of polysaccharides from Clematis Chinensis on the human tongue squamous cell carcinoma Tca-8113 cells.Methods:Conventional cultured human tongue squamous carcinoma cell Tca-8113,choose logarithmic growing phase cells as experiment subjects, according to the different concentration of drug, we could set five experiment groups in all(each group includes four parallel holes and test it again and again for three times), adjust the concentration of 1x104 cells / ml and seed it in 96-hole plate, then add some fresh medium into a blank controlled group, and add the clematis polysaccharides of 0, 50, 100, 200, 400, 500 ug/ml(final concentration) to the five experiment groups separately, keep training for 24hrs and determine OD value of each hole at 570nm wavelength on the enzyme-labeled reader, test the situation of polysaccharide on the growth inhibition of Tca-8113 at different concentration with the method of MTT; choose five experiment groups of clematis polysaccharide of final concentration 100ug/ml, train them separated after 12hs, 24hs, 48hs, 72hs, 96hs, use the method of MTT to observe the situation of clematis polysaccharide on the growth inhibition of Tca-8113 at a certain concentration and different time; adjust the concentration of 1x106 cells / ml and seed it in six-hole plate, add some fresh medium into a blank controlled group, set five experiment groups again(test it repeatedly for three times), clematis polysaccharide of final concentration 100ug/ml is positive controlled group, reflect on cells Tca-8113, train them separated after five days, use the staining method PI to test the change of cells cycle, and the Annexin V-FITC/PI double staining method, cytometry to determine apoptosis of cells, finally use statistics to analysis the results of experiments.Results: All different concentration of clematis polysaccharide has a clear inhibitory effect on the growing of Tca-8113 cells, as the clematis polysaccharide's concentration and time increases, the inhibitory effect also gradually increased, it shows a dependent relationship of certain dose and time, and the effect of Tca-8113 cells growing inhibitory of the group of high concentration of clematis polysaccharide has a meaning of Statistics; apply cytometry on determining that, clematis polysaccharide has different effects on various stages of Tca-8113 cell cycle, the cells'proportion of stage G1 increases, the cells'proportion of stage S reduces, and has a dependence of time; the apoptosis rate of Tca-8113 cells become higher as the time increases, has a obvious meaning of statistics.Conclusion: 1. Clematis polysaccharide has a clear inhibitory effect on the growing of human tongue squamous carcinoma cell Tca-8113, and has a dependence of time and concentration. 2. Clematis polysaccharide reflects on the Tca-8113 cells'cycle, has a dependence of time, the difference has a meaning of statistics. 3. Clematis polysaccharide could lead Tca-8113 cells to the death, has a dependence of concentration and a meaning of statistics. 4. Use clematis polysaccharide alone or with other drugs may be a new kind of economical, effective and safe anti-tumor drug in clinical.