摘要
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。
Objective： To explore whether norcantharidin （NCTD） can enhance the cytotoxicity of IL-15 activated peripheral blood mononuclear cells （PBMCs） on human acute myeloblastic leukemic KG1a cells and its underlying mechanism. Methods： The effect of NCTD on the proliferation of KG1a cells was detected by typan blue assay and CCK-8 assay. The effect of NCTD on the cell cycle of KG1a cells was examined by flow cytometry. The cytotoxicity of IL-15 activated PBMCs （IL-15-PBMCs） against NCTD treated-KG1a cells was detected by LDH releasing assay. The expressions of NKG2D （natural killer group 2 member D） ligands on KG1a cells were detected by flow cytometry. Results： NCTD effectively inhibited the proliferation of leukemic KG1a cells, in a time- （r=0.398，P=0.000） and dose-dependent manner （r=0.861，P=0.000）, and arrested KG1a cell cycle at G2/M phase. NCTD within a concentration of 4.00 μg/ml has no obvious cytotoxicity on the IL-15 activated PBMCs （IL-15-PBMCs） （P〉0.05）. Compared with the control group, the cytotoxic rate of IL-15-PBMCs on 0.125 μg/ml NCTD treated-KG1a cells was significantly increased （donor A： ／[37.44±5.78／]% vs ／[9.33±1.69／]%, ／[38.33±3.07／]% vs ／[16.75±1.20／]%, P〈0.05）. NCTD treatment showed no effect on expressions level of NKG2D ligands on KG1a cell surface （P〉0.05）. Conclusion： NCTD can enhance the cytotoxicity of IL-15-PBMCs on leukemic KG1a cells, which is possibly related to the inhibition of proliferation of KG1a cells and cell cycle arrest in G2/M phase.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2013年第5期580-585,共6页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金资助项目(No.30973454)
教育部新世纪优秀人才支持计划资助项目(No.NCET-09-0087)~~
