Effects Of SCD40L On Proliferation And Apoptosis Of Leukemia Jurkat Cell Line

Objective: To study the effect of s CD40L(soluble CD40 ligand)on the proliferation and apoptosis of Jurkat cell line in vitro,and to provide experimental basis for the clinical treatment of leukemia.Methods :(1)the effects of different concentrations of s CD40 L on the proliferation inhibition rate of Jurkat cells(Acute T lymphoblastic leukemia cells)cultured in vitro were detected by CCK8 method,and the optimal subsequent concentration was determined.(2)Flow cytometry(Annexin V-FITC/PI double staining)was used to determine the effect of s CD40 L at the optimal concentration on Jurkat cell apoptosis.(3)The effects of s CD40 L on the m RNA expression levels of apoptosis-related genes Bcl-2 and Bax in Jurkat cells were detected by real-time fluorescence quantitative PCR(RT-PCR).Results:(1)CCK8 method showed that: Add s CD40 L with different concentrations(1.00μg/ml,2.00 μg/ml,3.00 μg/ml,4.00 μg/ml,5.00 μg/ml,6.00 μg/ml)at the same time,After incubating with Jurkat cells for 24 h,48 h and 72 h,they had different degrees of proliferation inhibition.Compared with blank control group and daunorubicin group(positive control group),the proliferation inhibition rates were statistically significant(P<0.05).s CD40 L with a concentration of 5.00 μg/ml had the most obvious inhibitory effect on the proliferation of Jurkat cells,so 5.00 μg/ml was selected as the subsequent experimental concentration.Under the same concentration and different action time,s CD40 L and Jurakt cells incubated for 72 h had the most significant inhibitory effect on the proliferation of Jurkat cells.(2)Apoptosis was detected by flow cytometry(Annexin V-FITC/PI double staining)and the results showed that: s CD40 L with drug concentration of 5.00 μg/ml was incubated with Jurkat cells for 24 h,48 h and 72 h,and there were statistically significant differences in apoptosis rate compared with blank control group and daunorubicin group(positive control group)(P<0.05).s CD40 L had the most significant apoptotic effect on Jurkat cells at 48 h under the same concentration and different treatment time.(3)RT-PCR detection results showed that: After treatment with s CD40 L at5.00 μg/ml for 24 h,48 h and 72 h,the expression level of Bcl-2 m RNA decreased compared with the blank control group,with statistical significance(P<0.05),and the decrease degree was most significant at 48 h.Compared with blank control group,the expression level of Bax m RNA was increased,the differences were statistically significant(P<0.05),and the increase degree was most significant at 48 h.Conclusion :(1)s CD40 L can inhibit the proliferation of Jurkat cells in vitro and promote the apoptosis of Jurkat cells.(2)s CD40 L has certain anti leukemia effect on Jurkat cells in vitro,and the mechanism may be related to the participation of Bcl-2 / Bax pathway.