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Objective: strict vitro study ginseng polysaccharides (GPS)-induced HL-60 (acute promyelocytic leukemia cell line) during apoptosis survivin and caspase-3 changes. Understanding of different concentrations of ginseng polysaccharides on HL-60 proliferation, apoptosis, and the induction of apoptosis in HL-60 concentration. Expect to find a new drug can be used alone or in combination with other drugs to relieve the patient's symptoms, improve the quality of life of patients. Methods: 1. RPMI1640 medium containing 5% fetal bovine serum in cultured human leukemia cell line HL-60 in 25ml of culture flasks, placed in 37 ° C, 5? Saturated humidity incubator culture, every 2 to passaged three days, taking the logarithmic growth phase cells experiments. The cell concentration was adjusted to 2 × 103/ml cells were seeded in 96-well microtiter plates, and a total of five experimental groups (n = 3 parallel empty, and repeat the experiment) the cell suspension was added to each well 50ul, the final concentration of containing the GPS 50μg/ml, 100μg/ml, 150 μg, 200μg/ml, 250μg/ml each 50ul for the experimental group, blank group with equal amount of media, the role of 24h, 48h respectively after 72 hours by CCK -8 kit detects GPS inhibition of HL-60 and the inhibition rate was calculated. The cell concentration was adjusted to 5 × 102/ml cell suspension was seeded in six-well plates, each well plus the cell suspension 2ml first hole add 2ml medium for the negative group, the remaining five holes were added in a final concentration of 50μg / ml and 100μg/ml 150μg / ml, 200μg/ml, 250μg/ml of GPS 2ml experimental group for 72h after the enzyme-linked immunosorbent assay (ELISA) to detect the expression of apoptosis inhibitory factor (Survivin). 4 and the cell concentration was adjusted to 2 × 106/ml cell suspension was seeded in six-well plates, and added to each well of cell suspension 1ml, first holes add 1ml of culture medium as a negative group, the remaining five holes were added in a final concentration of 50μg / ml, 100μg/ml, 150 μg, 200μg/ml, 250μg/ml's GPSlml of the experimental group, caspase-3 activity for 72h with enzyme-linked immunosorbent assay (ELISA) detection of apoptosis-related factors. Results: 1.CCK-8 results show that different concentrations of GPS (50μg/ml, 100μg/ml, 150 μg, 200μg/ml, 250μg/ml) on HL-60 are inhibited, and was time-and concentration-dependent manner With GPS concentration increased and prolonged duration of action, inhibition enhanced. Conclusion: 1. GPS HL-60 leukemia cells to significantly inhibited appreciation was time concentration-dependent. 2 enzyme-linked immunosorbent assay (ELISA) detection of inhibitors of apoptosis (Survivin) results showed that different concentrations of ginseng polysaccharides (50μg/ml, 100μg/ml, 150 μg, 200μg/ml, 250μg/ml) reversed apoptosis inhibition The factor (Survivin) inhibited with ginseng polysaccharide concentration increased apoptosis inhibiting factor Survivin reduce. 3. Caspase family is a key component in the process of apoptosis, activation or abnormal expression caused apoptosis, also known as the death protease, can interact with numerous protein factors regulate cell apoptosis. 14 family members has been found that the caspase-3 is the most important effect type caspase. Most of the factors that trigger apoptosis, and eventually go through caspase-3-mediated signal transduction pathways leading to apoptosis. The enzyme-linked immunosorbent assay (ELISA) test results showed that different concentrations of ginseng polysaccharides (50μg/ml, 100μg/ml, 150μg/ml, 200μg/ml, 250μg/ml) can activate apoptotic factor caspase-3, by caspase-3-mediated signal transduction pathways leading to apoptosis of HL-60 cells, caspase-3 activity increased with the concentration of ginseng polysaccharides. 4.GPS an important role in the clinical treatment of leukemia, in combination with other chemotherapy drugs may achieve better results.
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