Inhibition Mechanism Research About CD59 Specific Site Closed Short Peptide On T Cell Leukemia

Objective To study the effect of CD59 specific active site closed Short peptide on Jurkat cell signal transduction molecules(Lat,Stat3,Nf-Kb3,BcL-xL,Caspase-3),research the specific mechanism that short peptide of CD59 specific site closed SP22 action on Jurkat cells.Build the T department Jurkat cell leukemia nude mice model,study the role of closed short peptide SP22 on Jurkat cell transmembrane signal transduction of nude mice leukemia by CD59 mediated,to provide experimental basis for the research and treatment of clinical T department leukemia.Methods One,Cells Experiment:cells divided into two group,?Jurkat cells control group(group Jurkat)?add SP22 ligand peptide Jurkat cells group(SP22 group).Training after 48h respectively,using CCK and BRDU method to detect cell proliferation rate;using Flow cytometry instrument apoptosis of two groups cells;Real time quantitative PCR to detect the expression of joint protein Lat and gene Stat3,Nf-?b3,BcL-xL,Caspas-3 before and after the experiment of two groups from genes level,ELISA detection related cytokines;Western blot detection related proteins expressed in the signal path.Two,Animal experiment:build Jurkat cell leukemia nude mouse model:24 BALB/c-nu nude mice were divided into three group randomly,each group have 8.Set?injection PBS group(blank group);?injection Jurkat cells group(group Jurkat)?injection via SP22 ligand peptide closed Jurkat cells group(group SP22);Three groups mice were injected 4 weeks,The mice were weighed respectively in 0,1,2,3,4week before and after injection.The white blood cell number in peripheral blood were counted in 0,1,2,3,4weeks.flow cytometry instrument detection apoptosis in Jurkat cells in bone marrow and peripheral blood;ELISA test mice serum IL-2 and TNF-alevel;The mice liver,spleen tissue slice HE dyeing observation pathological changes;Immunohistochemical detection the expression of the BcL-xL and the Caspase-3.in mice liver and spleen.Results One,Cells Experiment SP22 cell proliferation inhibition rate is greater than the Jurkat group(P<0.05);SP22 group compared with Jurkat group found drop in the gene expression of Lat,Stat3,Nf-Kb3 and BcL-xL(P<0.05),gene expression of Caspas-3 is increased(P<0.05);SP22 group add concentration of 40mg/ml group increased significantly in apoptosis ratio,cell DNA synthesis period ratio decreased significantly,compared with Jurkat group differences significant(P<0.05);SP22 group compared with Jurkat group,cell secretion and promote tumor cell growth of interleukin 2(IL-2)and TGF-? significantly decreased(P<0.05).Two,Animal experiment after the success of the nude mouse leukemia model building,SP22 group mice individual quality is lower than the blank group,but more than Jurkat group.SP22 group mice peripheral blood leukocyte count more than the blank group,less than Jurkat group(P<0.05).Flow cytometry instrument detect the bone marrow and peripheral blood Jurkat cells apoptosis level of mice showed SP22 higher than Jurkat group(P<0.05).ELISA test showed that SP22 group mice serum IL-2 and TNF-alevel lower than the Jurkat group(P<0.05).Tissue section HE staining showed that liver tissue of mice blank group is normal,Jurkat group and SP22 group all can saw the Jurkat cells infiltration,disorganized and occasional false flocculus to form.Jurkat group the Jurkat cells infiltrating is greater than SP22 group.blank group of mice spleen tissue is no problem,Jurkat group and SP22 group of mice spleen tissues have Jurkat cells infiltration,visible structure disorder of different level,Jurkat group the Jurkat cells infiltrating is greater than SP22 group.Immunohistochemical display the expression level of Caspase 3 in the liver and spleen of mice that the SP22 group higher than Jurkat group.the BcL-xL expression level in the liver and spleen of mice show that the SP22 group is lower than the Jurkat group.Conclusion Through cells and animal experiments have verified that closed short peptide SP22 of the specific site CD59 have inhibit proliferation effection on Jurkat cells of T department leukemia,reduce the expression of Lat,Stat3,Nf-Kb and BcL-xL signaling molecules,promote the activation of caspase-3 and promote apoptosis of Jurkat cells.This study not only provide the experimental basis for the role of T department leukemia cell signal transduction of short closed peptide at CD59,but also shows a broad prospect.for the targeted therapy of acute T lymphocyte leukemia.