Objective To study the effect of γ-interferon （IFNγ）, tumor necrosis factor related apoptosis inducing ligand （TRAIL）, and cisplatin or etoposide induced apoptosis in human neuroblastoma cell line SH-SY5Y and it...Objective To study the effect of γ-interferon （IFNγ）, tumor necrosis factor related apoptosis inducing ligand （TRAIL）, and cisplatin or etoposide induced apoptosis in human neuroblastoma cell line SH-SY5Y and its possible molecular mechanisms. Methods The expressions of Caspase 8 mRNA and protein were detected with RT-PCR and Western blot analysis. The effects of IFNγ, TRAIL, IFNγ ＋ TRAIL, IFNγ ＋ Caspase 8 inhibitor ＋ TRAIL, IFNγ ＋ cisplatin ＋ TRAIL, and IFNγ ＋ etoposide ＋ TRAIL on the growth and apoptosis of SH-SY5Y cells were detected with the methods of MTT and flow cytometry. The relative Caspase 8 activity was measured with colorimetric assay. Results Caspase 8 was undetectable in SH-SY5Y cells but an increased expression of Caspase 8 mRNA and protein was found after treatment with IFNγ. SH-SY5Y ceils themselves were not sensitive to TRAIL, but those expressing Caspase 8 after treatment with IFNγ were. The killing effect of TRAIL on SH-SY5Y cells expressing Caspase 8 was depressed by Caspase 8 inhibitor. Cisplatin and etoposide could enhance the sensitivity of TRAIL on SH-SY5Y cells. The relative Caspase 8 activity of SH-SY5Y cells in IFNγ ＋ TRAIL group was significantly higher than those of control group, IFNγ group, TRAIL group, and inhibitor group （ P 〈 0. 01 ）. There was no significant difference among IFNγ ＋ TRAIL group, IFNγ ＋ cisplatin ＋ TRAIL group, and IFNγ ＋ etoposide ＋ TRAIL group. Conclusions IFNγ could sensitize SH-SY5Y cells to TRAIL-induced apoptosis and this may be realized by the up-regulation of Caspase 8. Cisplatin and etoposide could enhance the killing effect of TRAIL on SH-SY5Y cells.展开更多
Objective To study the changes of lymphocyte subsets and regulatory T cells in peripheral blood of patients with acute leukemia(AL) and its clinical significance.Methods The different levels of peripheral blood lympho...Objective To study the changes of lymphocyte subsets and regulatory T cells in peripheral blood of patients with acute leukemia(AL) and its clinical significance.Methods The different levels of peripheral blood lymphocyte subsets and regulatory T cells of 60 AL patients and 40 normal controls were detected with flow cytometry.Results Compared with the normal controls,the percentages of CD3+ T cells,CD4+ T cells,CD16+CD56+ NK cells and the ratio of CD4+ /CD8+ obviously decreased in newly diagnosed AL group(P <0.05),while their percentages of CD8+ T cells and CD19+ B cells significantly increased(P <0.01).The percentage of CD4+ T cells and the ratio of CD4+ /CD8+ in acute lymphoblastic leukemia(ALL) group were much lower than those in acute myelogenous leukemia(AML) group(P <0.01).Compared with these in control group,the proportions of CD4+ CD25high Treg cells and CD4+ CD25+ T cells in newly diagnosed AL group were significantly increased(P <0.01).Conclusion Cellular immune function is significantly abnormal in patients with AL.Compared with AML patients,ALL patients had poorer cellular immune function.The increased CD4 + CD25high Treg cells might be one of the important reasons of immunosuppression in AL.Detection of lymphocyte subsets and regulatory T cells is of clinical value on the evaluation of therapeutic effect and prognosis in AL patients.展开更多
基金the National Natural Sciences Foundation of China(39470739)the Ministry of Public Health Research Foundation(20122167)the Doctor Startup-Natural Science Foundation of Li-aoning Province (20041047)
文摘Objective To study the effect of γ-interferon （IFNγ）, tumor necrosis factor related apoptosis inducing ligand （TRAIL）, and cisplatin or etoposide induced apoptosis in human neuroblastoma cell line SH-SY5Y and its possible molecular mechanisms. Methods The expressions of Caspase 8 mRNA and protein were detected with RT-PCR and Western blot analysis. The effects of IFNγ, TRAIL, IFNγ ＋ TRAIL, IFNγ ＋ Caspase 8 inhibitor ＋ TRAIL, IFNγ ＋ cisplatin ＋ TRAIL, and IFNγ ＋ etoposide ＋ TRAIL on the growth and apoptosis of SH-SY5Y cells were detected with the methods of MTT and flow cytometry. The relative Caspase 8 activity was measured with colorimetric assay. Results Caspase 8 was undetectable in SH-SY5Y cells but an increased expression of Caspase 8 mRNA and protein was found after treatment with IFNγ. SH-SY5Y ceils themselves were not sensitive to TRAIL, but those expressing Caspase 8 after treatment with IFNγ were. The killing effect of TRAIL on SH-SY5Y cells expressing Caspase 8 was depressed by Caspase 8 inhibitor. Cisplatin and etoposide could enhance the sensitivity of TRAIL on SH-SY5Y cells. The relative Caspase 8 activity of SH-SY5Y cells in IFNγ ＋ TRAIL group was significantly higher than those of control group, IFNγ group, TRAIL group, and inhibitor group （ P 〈 0. 01 ）. There was no significant difference among IFNγ ＋ TRAIL group, IFNγ ＋ cisplatin ＋ TRAIL group, and IFNγ ＋ etoposide ＋ TRAIL group. Conclusions IFNγ could sensitize SH-SY5Y cells to TRAIL-induced apoptosis and this may be realized by the up-regulation of Caspase 8. Cisplatin and etoposide could enhance the killing effect of TRAIL on SH-SY5Y cells.
文摘Objective To study the changes of lymphocyte subsets and regulatory T cells in peripheral blood of patients with acute leukemia(AL) and its clinical significance.Methods The different levels of peripheral blood lymphocyte subsets and regulatory T cells of 60 AL patients and 40 normal controls were detected with flow cytometry.Results Compared with the normal controls,the percentages of CD3+ T cells,CD4+ T cells,CD16+CD56+ NK cells and the ratio of CD4+ /CD8+ obviously decreased in newly diagnosed AL group(P <0.05),while their percentages of CD8+ T cells and CD19+ B cells significantly increased(P <0.01).The percentage of CD4+ T cells and the ratio of CD4+ /CD8+ in acute lymphoblastic leukemia(ALL) group were much lower than those in acute myelogenous leukemia(AML) group(P <0.01).Compared with these in control group,the proportions of CD4+ CD25high Treg cells and CD4+ CD25+ T cells in newly diagnosed AL group were significantly increased(P <0.01).Conclusion Cellular immune function is significantly abnormal in patients with AL.Compared with AML patients,ALL patients had poorer cellular immune function.The increased CD4 + CD25high Treg cells might be one of the important reasons of immunosuppression in AL.Detection of lymphocyte subsets and regulatory T cells is of clinical value on the evaluation of therapeutic effect and prognosis in AL patients.
