斑蝥酸钠对膀胱癌患者树突状细胞及BIU-87细胞的影响  

Effects of sodium cantharidinate on dendritic cells in patients with bladder carcinoma and BIU-87 cells

作　　者：宁波 李朵璐[1] 阚全程[1] NING Bo;LI Duolu;KAN Quancheng(First Affiliated Hospital,Zhengzhou University,Zhengzhou,Henan 450000,China)

机构地区：[1]郑州大学第一附属医院肿瘤科,450000

出　　处：《重庆医学》2018年第27期3494-3498,共5页Chongqing medicine

摘　　要：目的探讨斑蝥酸钠对膀胱癌患者外周血来源的树突状细胞(DC)成熟、功能的影响及斑蝥酸钠对膀胱癌细胞BIU-87增殖、凋亡的影响。方法分离15例临床确诊的中晚期膀胱癌患者外周血单核细胞,在含粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素-4(IL-4)及不同浓度斑蝥酸钠培养条件下制备DC。观察DC形态学变化,流式细胞仪检测DC表面CD1a和CD83表达水平,并用MTT法测定DC刺激同种异体淋巴细胞增殖能力。采用MTT法测定不同浓度斑蝥酸钠作用后BIU-87细胞的生长情况。Annexin V-FITC/PI双染流式细胞术观察斑蝥酸钠作用后细胞的凋亡情况。Western blot法检测Caspase-3和PARP的表达的变化。结果在斑蝥酸钠高剂量组,DC膜表面分子CD83、CD1a的表达最高。与对照组相比,斑蝥酸钠处理组DC膜表面分子CD83和CD1a表达增高(P<0.05)。斑蝥酸钠处理组刺激异体淋巴细胞增殖指数显著高于对照组。MTT法证实斑蝥酸钠能明显抑制膀胱癌细胞株BIU-87的生长。斑蝥酸钠作用24h后,流式细胞术双染法检测细胞凋亡显示斑蝥酸钠可使BIU-87细胞发生凋亡;Western blot法检测显示斑蝥酸钠使BIU-87细胞中Caspase-3和PARP蛋白表达明显增加。结论斑蝥酸钠能够诱导膀胱癌患者DC成熟,促进淋巴细胞的增殖能力,斑蝥酸钠能够抑制膀胱癌BIU-87细胞的增殖,诱导细胞凋亡。Objective To investigate the effect of sodium cantharidinate on the maturation and function of peripheral blood monocyte-derived dendritic cells(DC)from bladder carcinoma patients and to study its effect on the proliferation and apoptosis of human bladder cancer BIU-87 cells.Methods The peripheral blood monoeyes in the patients with middle and advanced stage bladder carcinoma were separated for preparing DC under the culture condition containing granuloeyte-macrophage colony-stimulating factor(GM-CSF)plus IL-4 and different concentrations of sodium cantharidinate.The cell morphology was observed and DC surface CD1a and CD83 expression levels were detected by flow cytometry.Allogeneic lymphocytes proliferation ability stimulated by DC was detected by methyl thiazolyl tetrazolium(MTT).The cell growth situation of BIU-87 cells after action by different concentrations of sodium cantharidinate was detected by MTT assay.The AV-PI double staining flow cytometry was carried out to detect the cell apoptosis.And the Caspase-3 and PARP expression changes were detected by Western blotting.Results The expression levels of DC phenotype molecule CD83 and CD1a in the sodium cantharidinate high dose group were the highest.Compared with the control group,DC membrane molecule CD83 and CD1a expression in the sodium cantharidinate treatment group was increased(P<0.05).The stimulating allogeneic lymphocytes proliferation index in the sodium cantharidinate treatment group was significantly higher than that in the control group.The MTT method verified that sodium cantharidinate could effectively inhibit the growth of bladder cancer BIU-87 cells.After 24 h action by sodium cantharidinate treatment,detecting cellular apoptosis by flow cytometry double-staining method showed that sodium cantharidinate could make the BIU-87 cells apoptosis,and Western blot detection showed that sodium cantharidinate could make the expression of Caspase-3 and PARP proteins in BIU-87 cells was markedly increased.Conclusion Sodium cantharidinate can induce the

关 键 词：树突状细胞 膀胱肿瘤 斑蝥酸钠 凋亡 

分 类 号：R737.14[医药卫生—肿瘤]