Cbp Overexpression And Palmitoylation On CD59 - Mediated Transduction Of Transmembrane Signal Transduction In Jurkat Cells Of T - Line Leukemia In Nude Mice

Objective To explore the role of transmembrane apoptotic signal transduction about over expression of Cbp and Cbp palmitoylation mutation to Jurkat cells of T lineage leukemia in nude mice.Methods Cbp-EGFP and Cbp-m-EGFP slow virus vector were constructed for establishing stable Jurkat cell lines. The transfection efficiency and location of Cbp and CD59 in Jurkat cells were researched by laser confocal microscope. The 32 female BALB/c nude mice were randomly distributed into blank control group, normal Jurkat cell control group(normal control group), over expression of Cbp model group(over expression of Cbp group) and Cbp palmitoylation mutation model group(Cbp-m group),then the mice were injeced 1×107/0.1 ml Jurkat cells. The mice were weighed in 0, 1, 2, 3,4 weeks. The white blood cell in peripheral blood were counted in 0, 1, 2, 3, 4 weeks. The apoptosis of Jurkat cells in peripheral blood were tested by laser confocal microscope.The apoptosis of Jurkat cells in the bone marrow and peripheral blood were detected with flow cytometry, the IL-2 concentration were detected using ELISA. The HE staining was used to observe the pathological changes of liver and spleen.The expression of Fas and survivin in liver and spleen were detected with immunohistochemical staining.Results The transfection efficiency of Jurkat cells was as high as 100%, Cbp and CD59 were located on the membrane of Jurkat cells. Compared with Cbp-m group, the weight of mice in over expression of Cbp group was higher(P<0.05), the amount of white blood cell in over expression of Cbp group was lower(P<0.05). Compared with Cbp-m group,the apoptotic level of Jurkat cells in bone marrow and peripheral blood in over expression of Cbp group were higher(P<0.05) with flow cytometry; the IL-2 concentration in over expression of Cbp group was lower(P<0.05) using ELISA. HE results showed that the structure of liver and spleen were disorder, hepatic lobule structure was disorder.Compared with Cbp-m group, the expression of Fas in over expression of Cbp group were higher(P<0.05), the expression of survivin in over expression of Cbp group were lower(P<0.05) with immunohistochemical staining.Conclusion This topic confirmed that CD59(GPI anchoring protein) transduced transmembrane apoptotic signal by Cbp in nude mice model with T lineage leukemia Jurkat cells. Over expression of Cbp can promote the apoptosis of T lineage leukemia Jurkat cells, the apoptotic efficiency of Jurkat cells was declined after Cbp palmitoylation mutation. The study provided a new mentality for Cbp and CD59 in the role of apoptotic signal transduction mechanism in T cell, and provided a new target for the therapy of acute T lineage leukemia.