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目的:探讨去甲斑蝥素(norcantharidin,NCTD)是否能增强IL-15活化的人外周血单个核细胞(peripheral blood mononuclear cell,PBMC)对人急性髓系白血病KG1a细胞的杀伤作用及其可能机制。方法:锥虫蓝拒染法、CCK-8法检测NCTD对KG1a细胞增殖的影响,流式细胞术检测NCTD对KG1a细胞周期的影响,LDH释放法检测IL-15活化的PBMC(IL-15-PBMC)对NCTD处理后KG1a细胞的细胞毒活性,流式细胞术检测KG1a细胞表面NKG2D(natural killer group 2 member D)配体的表达。结果:NCTD有效抑制白血病KG1a细胞的增殖,呈时间(r=0.398,P=0.000)和剂量依赖性(r=0.861,P=0.000),并阻滞KG1a细胞周期于G2/M期;4μg/ml以下的NCTD对IL-15-PBMC没有明显的增殖抑制作用(P>0.05)。当效靶比为10∶1和20∶1时,IL-15-PBMC对0.125μg/ml NCTD处理后KG1a细胞的杀伤率较对照组明显增加[志愿者A:(37.44±5.78)%vs(9.33±1.69)%,(38.33±3.07)%vs(16.75±1.20)%;P<0.05]。NCTD不影响KG1a细胞表面NKG2D配体蛋白的表达(P>0.05)。结论:NCTD能增强IL-15-PBMC对白血病KG1a细胞的杀伤作用,可能与抑制细胞增殖、阻滞细胞周期于G2/M期有关。
AIM: To investigate whether norcantharidin (NCTD) can enhance the cytotoxicity of human peripheral blood mononuclear cells (PBMC) activated by IL-15 on human acute myeloid leukemia cell line KG1a and its possible mechanism. Methods: The effects of NCTD on the proliferation of KG1a cells were detected by CCK-8 assay, the cell cycle of KG1a cells was detected by flow cytometry, the expression of IL-15-activated PBMCs was detected by LDH release assay, PBMC) on KG1a cells after NCTD treatment, and the expression of NKG2D (natural killer group 2 member D) ligand on KG1a cells was detected by flow cytometry. Results: NCTD effectively inhibited the proliferation of KG1a leukemia cells in a dose-dependent manner (r = 0.398, P = 0.000) and in a dose-dependent manner (r = 0.861, ml NCTD IL-15-PBMC no significant inhibition of proliferation (P> 0.05). IL-15-PBMC significantly increased the killing rate of KG1a cells treated with 0.125μg / ml NCTD compared with the control group when the target ratios were 10:1 and 20:1 [Vol A (37.44 ± 5.78)% vs 9.33 ± 1.69%, (38.33 ± 3.07)% vs (16.75 ± 1.20)%, respectively; P <0.05]. NCTD did not affect KG1a cell surface NKG2D ligand protein expression (P> 0.05). CONCLUSION: NCTD can enhance the cytotoxicity of IL-15-PBMC on KG1a leukemia cells, which may be related to inhibiting cell proliferation and arresting cell cycle in G2 / M phase.