Study On The Induction Of Apoptosis In Human Promyelocytic Leukemia Cell By Ginseng Polysacchride

By rigorous experiment in Vitro, study on the change of survivin and capase-3 in the process of apoptosis in HL-60 inducted by GPS. To know the different concentrations of GPS in effects on the Proliferation and Apoptosis in HL-60 Cells, and the best concentrations of GPS that induct HL-60. To expect a new medicine which can alone or in combination with other medicines to lighten patients's symptom, improve the general condition. Methods:1. HL-60 was cultured with RPMI1640 containing 5%fetal bovine serum in 25ml culture flask, in the educated box of 5%CO2,saturated degree of humidity, in 37℃, harvest every3～4days, cells in logarithmic phase are used in the experiment.2. Adjust the cell concentration to 2×103 cells/ml and incubate to 96 hole cell culture flask, divided into 5 groups(3 replicates pretreatment), each well contained 50ul, take 50ul uptitratal concentrations of GPS (50μg /ml,100μg/ml,150μg/ml,200μg/ml,250μg/ml) as the experimental groups, add same amount culture medium to the model group, the inhibition of GPS on HL-60 is detected by CCK-8 kit at the 24h,48h,72h, and the inhibitory rate is calculated.3. Adjust the cell concentration to 5×102 cells/ml and incubate to 6 hole plate, each well contained 2ml, add 2ml culture medium to the first well as negative group, add 2ml uptitratal concentrations of GPS (50μg/ml,100μg/ml,150μg/ml,200μg/ml,250μg/ml) to other 5 wells as experimental groups, the expression of Survivin is detected by ELISA after 72h.4. Adjust the cell concentration to 2×106 cells/ml and incubate to 6 hole plate, each well contained 1ml, add 1ml culture medium to the first well as negative group, add lml uptitratal concentrations of GPS (50μg/ml,100μg/ml,150μg/ml,200μg/ml,250μg/ml) to other 5 wells as experimental groups, the activity of caspase-3 is detected by ELISA after 72h. Results:1. Results of CCK-8 display different concentrations of GPS (50μg/ml,100μg/ml,150μg/ml,200μg/ml,250μg/ml) have inhibited growth of HL-60, and in concentration and time dependent manners, the inhibitory effect increased with the increase of GPS's concentration and time. Conclusion:1. GPS has significant inhibitory effects on HL-60, in concentration and time dependent manners.2. Results of Survivin detected by ELISA display different concentrations of GPS (50μg/ml,100μg/ml,150μg/ml,200μg/ml,250μg/ml) have inhibited growth, Survivin decrease with the increase of GPS's concentration.3. Pan-caspase acting element in the process of apoptosis, activation and exceptionally expression is capable of causing apoptosis, is also called death protease, interaction of multitudinous proteins regulate cell apoptosis.14 members have heen identified, caspase-3 is the most important responsive caspase. Majority trigger apoptosis factors, all ultimately end up using transmit signal mediated by caspase-3 results in cell apoptosis. Results detected by ELISA display different concentrations of GPS(50μg/ml,100μg/ml,150μg/ml,200μg/ml,250μg/ml) active apoptosis related proteins, use transmit signal mediated by caspase-3 results in HL-60 cell apoptosis, activity of caspase-3 enhance with the increase of GPS's concentration.4. There is an important action of GPS to treat leukaemia, combine with other medicines can reach more ideal result.