The Effects Of DADS On Proliferration And Invasion Of Lymphoma Cells By Down Regulate DJ-1 Through Blocking SRC And ERK Pathway

[Objective] To detect the expression of DJ-1 protein in tissue of malignant lymphoma, reactive lymphoid hyperplasia and normal lymph node, study the effect of Diallyl Disulfide(DADS) on DJ-1expression and Src, Erk signaling pathway and biological behavior of the Raji and OCI-LY1 cells, sought to providing new clues to treat lymphoma.[Methods] By tissue microarray and immunohistochemical investigate DJ-1 protein expression of different types of lymphoma,reactive lymphoid hyperplasia and normal lymph node tissue; Treated Burkitt lymphoma Raji cells and diffuse large B cell lymphoma OCI-LY1 cells with different concentrations of DADS, by CCK 8 to assess proliferation inhibition; Flow cytometry detect cell cycle; Giemsa dyeing experiment detect cell morphology changes; Transwell experiment test DADS, Src inhibitor and Erk inhibitor effects on cell migration and invasion ability; Western blot to investigate the effects of DADS on DJ-1,p-Src, p-Erk expression of lymphoma cells, as well as the effects of Src inhibitor, Erk inhibitor. [Results] Tissue microarray results showed that the high expression rate of nucleus DJ-1 in Hodgkin lymphoma and non Hodgkin lymphoma cell were 42.9% and 72.1%, significantly higher than that in non tumor tissue 14.3%(reactive lymphoid hyperplasia and normal lymph node tissue)(P<0.05), high expression rate of plasma was100%, 93.4%, significantly higher than the non tumor tissues of 28.6%( P<0.05). CCK-8 assay results showed that after different concentrations(2.5mg/L, 5mg/L, 10mg/L, 20mg/L, 40mg/L) DADS treat Raji cells 24 h, 48 h, 72 h, the protein absorbance value decreased, the proliferation inhibitory effect of DADS on Raji cells showed a time and concentration dependent. After different concentrations(5mg/L, 10mg/L,20mg/L, 40mg/L, 80mg/L) DADS treat OCI-LY1 cells 24 h, 48 h, 72 h, we get the similar results as in Raji cells. Flow cytometry showed that DADS increased G1 phase percentage of Raji cells and OCI-LY1 cells from54.43%, 51.89% to 47.72%,30.90% respectively(P<0.05), induced G1 phase cell block. Giemsa staining results showed that DADS can induce Raji cells and OCI-LY1 cells differentiate into normal lymph cells.Western blot results showed that: DJ-1 protein of Raji cells and OCI-LY1 cells showed a time-dependent decrease after DADS treat 12 h, 24 h, 48 h,(P<0.05), and the expression of p-Src and p-Erk significantly decreased after DADS treated 5min, 15 min, 30 min, 60 min( P<0.05), total expression of Src and Erk has no significantly change. After Src inhibitor,Erk inhibitor as well as their combination with DADS treat Raji cells and OCI-LY1 cells 12 h, 24 h, 48 h, DJ-1 protein levels showed a time-dependent decreased(P<0.05), and the levels of DJ-1 protein in the combined treatment group were down more significantly compared to the corresponding Src inhibitor, Erk inhibitor treatment group. Transwell assay showed that SRC inhibitor, ERK inhibitor cooperate with DADS decrease migration and invasion of Raji cells and OCI-LY1 cells.[Conclusion] 1.DJ-1 is abundantly expressed in the nucleus and cytoplasm of different lymphoma cells. 2.DADS inhibit the proliferation of lymphoma Raji and OCI-LY1 cells, reduce the migration and invasion ability by down-regulating DJ-1 protein through suppress Src, Erk signaling pathway.