The Effects Of Soluble CD40 Ligand On Human Leukemia Cell Line THP-1 Proliferation And The Expression Of PI3K、Akt MRNA

Objective:To observe the effect of s CD40 L on the cell proliferation, cell apoptosis, expression of PI3 K and Akt of acute monocytic leukemia(M5)cell line THP-1 cells.Methods:(1)THP-1 cells were used in this study,were co-incubated with sCD40 L of different concentrations. CCK-8 assay was used to detect different concentrations of s CD40 L effect on cells proliferation in vitro and chose the best concentration of s CD40 L.(2)THP-1 cell apoptosis rate was determined with Annexin V-FITC/PI double staining by flow cytometry.(3) RT-PCR was used to determine PI3 K and Akt m RNA expression.Results:(1)Compare with THP-1 cell groups,sCD40 L were able to inhibit THP-1 cells proliferation in 24、48、72 h(p <0.05),The decrease of growth inhibitory rate was most significant at its concentration of 4μg/ml. s CD40 L were co-incubated with on THP-1 cells for 24 h 、48h 、72 h,all had different degree of inhibition proliferation in vitro and s CD40 L 4μg/ml was the strongest at the 24h(p <0.05).(2)s CD40 L effected on THP-1 cells for three different time can be a significant apoptosis,compared with cells group(p <0.05), The apoptosis rate was most significant for 24 h.(3) s CD40 L 4ug/ml group compare with the control group for the same time,the expression of PI3 K and Akt m RNA was down-regulated(p <0.05), with the increase of time PI3 K and Akt m RNA up-regulated.Conclusion:(1) In vitro,4ug/ml sCD40 L induce THP-1 cells growth inhibition and apoptosis was most significant,the highest inhibition rate and apoptosis rate of THP-1 cells induced by s CD40 L was at 24 h.(2) In the process of THP-1 cells proliferation and apoptosis,the PI3K/Akt signaling pathways plays an important role.In another words,the molecular mechanism of THP-1 cells growth inhibition and apoptosis induced by s CD40 L is related to PI3K/Akt signaling pathways.