卵巢癌抗原冲击DC在体外对SKOV3细胞作用的研究  

The effect of ovarian cancer antigen impacted DC on SKOV3 in vitro

作　　者：盛敏佳[1] 薛丽娟[2] 王立岩[1] 郝筱诗[1] 刘玉萍[1] SHENG Min jia XUE Li-juan WANG Li-yan et al(China Japan Union Hospital of Jilin University ,Changchun 130033,China The First Hospital of Jilin University)

机构地区：[1]吉林大学中日联谊医院,吉林长春130033 [2]吉林大学第一临床医院

出　　处：《中国实验诊断学》2016年第11期1811-1816,共6页Chinese Journal of Laboratory Diagnosis

摘　　要：目的探讨卵巢癌患者外周血和腹水中培养的树突细胞(DC)在体外激活淋巴细胞产生的肿瘤特异性细胞毒T淋巴细胞(CTL)作用,及其对卵巢癌SKOV3细胞的杀伤作用。方法分别培养卵巢癌患者外周血及腹水来源DC,进行细胞形态学观察及细胞表面分子表达检测;以卵巢癌冻融抗原致敏DC,检测两种来源DC刺激T淋巴细胞增殖的能力及对IL-2水平的影响;检测抗原负载DC与T细胞诱导CTL对卵巢癌细胞的杀伤作用。结果镜下显示卵巢癌患者外周血及腹水来源的DC形态学无明显差异,腹水来源DC成熟时间较外周血来源DC晚,二者表面分子表达的阳性率除CD83外无明显差异;卵巢癌冻融抗原负载DC上调其表面相关分化抗原表达,统计学差异具有显著性(P<0.05);经卵巢癌冻融抗原负载后的DC刺激T淋巴细胞的增殖,差异有显著性(P<0.05),但两种来源DC间刺激指数无统计学差异;抗原负载DC与无抗原负载DC相比,其上清中IL-12浓度增高,差异具有显著性(P<0.05);负载与不负载抗原的DC对卵巢癌细胞杀伤作用增强,与对照组相比具有显著性差异(P<0.05),随着效应细胞数量的增高,杀伤作用增强,加入IL-2增强其杀伤作用,但外周血及腹水来源DC间相应情况下对卵巢癌细胞的杀伤作用无统计学差异(P<0.05)。结论卵巢癌患者外周血及腹水来源的DC均具有诱导CTL作用,DC作为载体负载肿瘤抗原后可以激发T淋巴细胞形成CTL,在体外有效的杀伤卵巢癌SKOV3细胞。Objective To study the effect of tumor specific cytotoxic T lymphocyte（CTL）produced by the activation of lymphocytes from the peripheral blood and ascites of dendritic cells（DC）in ovarian cancer patients in vitro,and its killing role on ovarian cancer cell SKOV3.Methods Cultured DC from blood and ovarian cancer ascites,observed the cell morphology and detected the cell surface molecules expression;DC was pulsed by freeze-thaw ovarian cancer antigen,to detect T lymphocyte proliferation and IL-2levels stimulated by two sources of DC;To detect killing roll of antigen load DC and T cells induce CTL for ovarian cancer cells.Results Microscopic display of peripheral blood and ascites derived DC morphology has no significant difference,maturation time of ascites source DC is later than peripheral blood DC;The positive rate of surface molecules expression has no significant difference except CD83;ovarian cancer lysates DC raised its surface associated antigen expression,the difference was statistically significant（P〈0.05）;The proliferation of T lymphocytes stimulated by ovarian cancer cell lysates DC was high,the difference was significant（P〈0.05）,but two sources of DC stimulation index has no significant difference;compared DC with antigen loaded,the DC without antigen loaded has supernatant IL-12 concentration increased,the difference was significant（P〈0.05）;Without loaded DC and with loaded DC antigen ovarian cancer killing effect enhanced as compared with the control group with significant difference（P〈0.05）;with the increasing number of cells,the killing effect after added IL-2was enhanced,but the peripheral blood DC sources and ascites has no significant difference（P〈0.05）.Conclusion The DC from peripheral blood and ascites of ovarian cancer patients have the effect of inducing CTL.DC loaded tumor antigen as a carrier can stimulate T lymphocytes formed CTL,that has anti-ovarian cancer cell SKOV3 effect in vitro.

关 键 词：卵巢癌 树突细胞 细胞毒T淋巴细胞 白细胞介素-2 杀伤作用 

分 类 号：R737.31[医药卫生—肿瘤]