CD95-mediated apoptosis in Burkitt's lymphoma B-cells is associated with Pim-1 down-regulation

Abstract

B-cells of the high-grade non-Hodgkin lymphoma Burkitt's lymphoma (BL) overexpress survival oncoproteins, including the proviral integration site for Moloney murine leukaemia virus kinase (Pim)-1, and become apoptosis resistant. Activated death receptor CD95 after ligation with anti-CD95 monoclonal antibody (mAb) resulted in the regression of BL via induction of apoptosis, suggesting a decrease of survival protein expression. Here, CD95-mediated apoptotic pathways in BL B-cell lines (Raji and Daudi) following treatment with anti-CD95 mAb was investigated with the cause-and-effects on pim-1 gene expression, in comparison with leukemic cell line (K562) used as CD95-negative cells. Immunohistochemical staining for CD95 and Pim-1 was performed, and the effects of anti-CD95 mAb on apoptotic signalling using western blotting, on caspase activity and cell survival of BL B-cell and leukemic cell lines were determined. We showed that Raji cells expressed more CD95 receptors than Daudi cells. Half of each population underwent apoptosis accompanied by decreased cell viability after anti-CD95 mAb treatment. Distinct extrinsic and intrinsic CD95-mediated apoptotic pathways in Raji and Daudi cells were revealed by high caspase activity and mitochondrial outer membrane permeabilization, respectively. We observed decreased Pim-1 transcript and protein expression levels with increased heat-shock protein (Hsp)70 and decreased Hsp90 expression in anti-CD95 mAb-treated cells. Throughout the study, K562 cells did not undergo apoptosis upon anti-CD95 mAb treatment. Pim-1 knockdown following to stable transfection with plasmid vectors induced apoptosis and decreased viability of BL and K562 cells. Therefore, CD95-mediated apoptosis induces Pim-1 down-regulation in BL B-cells, but Pim-1 down-regulation cannot fully eradicate BL and leukaemia.

Expression of PIM‑1 in salivary gland adenoid cystic carcinoma: Association with tumor progression and patients' prognosis

“…This suggested that upregulation of PIM-1 may promote the disruption of cell proliferation and homeostasis to drive malignant aggression and lead to a poor outcome. Dysregulation of apoptosis is a vital feature of oncogenes and a number of previous studies have revealed that PIM-1 has an important role in apoptosis (18)(19)(20). The present study demonstrated that the apoptotic rate had a significant association with PIM-1 expression level in ACC patients' tissues.…”

Anti-Proliferative and Pro-Apoptotic Effects ofCalligonum comosum(L’Her.) Methanolic Extract in Human Triple-Negative MDA-MB-231 Breast Cancer Cells

“…After the protein extraction from the untreated and treated TNBC MDA-MB-231 cells, 35 pro-apoptotic and anti-apoptotic proteins were detected, using the Proteome Profiler Human Apoptosis Protein Array kit (Life Technologies) and analyzed, as described in. 34…”

“…After the protein extraction from the untreated and treated TNBC MDA-MB-231 cells, 35 pro-apoptotic and anti-apoptotic proteins were detected, using the Proteome Profiler Human Apoptosis Protein Array kit (Life Technologies) and analyzed, as described in. 34 From cell lysis to the separation, by molecular weight, of the extracted proteins on 12% SDS-PAGE gel electrophoresis, Western blot technology was done as described in. 35 Provided by Abcam (Cambridge, UK), mouse monoclonal antibody against pro-and active cleaved-caspase-3 [ABM1C12], rabbit monoclonal antibody against p21 [EPR3993] (1:1000) and mouse monoclonal antibody against GAPDH (1:2000), were used as the primary antibodies for the detection of the targeted proteins.…”

In vitro assessment of the efficiency of the PIM‑1 kinase pharmacological inhibitor as a potential treatment for Burkitt's lymphoma

“…4309155; Qiagen GmbH), using the 7900 Fast Real-Time PCR system (Thermo Fisher Scientific, Inc.). The primer pair sequences (Invitrogen; Thermo Fisher Scientific) were used as previously described ( 12 ). The thermocycling conditions were as follows: Heat denaturing at 95°C for 10 min, then by 40 cycles of denaturing at 95°C for 10 sec followed by annealing at 57°C for 20 sec and finally extension at 72°C for 30 sec.…”