顺铂诱导肿瘤裂解物致敏DC抗肿瘤作用研究

目的探讨用顺铂(DDP)诱导卵巢癌细胞株(SKOV 3)获得的肿瘤细胞裂解物以致敏树突状细胞(dendritic cells,DCs)诱导的CTL对肿瘤细胞的杀伤作用。方法分离健康者外周血单个核细胞,培养DC,分为三组。顺铂组:用顺铂诱导卵巢癌细胞株获得细胞裂解物致敏DC;冻融组:用冻融法获得细胞裂解物致敏DC;空白组:不做任何处理。成熟的DC与T细胞混合培养获得特异性细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)。流式细胞仪(flow cytometry,FCM)检测三组DC表型,CTL亚群比例,MTT检测CTL对SKOV3的杀伤效果。结果顺铂组DC的分子表面标志CD 86、CD 40表达分别为(76.35±3.99)%、(71.20±6.86)%,显著高于空白组(P<0.05),而与冻融组相比差异无统计学意义;两组DC诱导的特异性CTL,其亚群CD 8+T、CD 4+T比值[(80.42±4.61)%、(9.55±2.24)%]明显著于空白组(P<0.05);顺铂组诱导的CTL对SKOV3的杀伤率随着E/T比例(20∶1,40∶1,80∶1)的增加而逐步渐增高,杀伤率[(45.66±3.16)%、(50.16±3.44)%、(58.82±4.06)%]显著高于冻融组[(39.71±4.77)%、(47.00±4.02)%、(53.76±4.78)%]与空白组[(37.94±3.74)%、(41.11±5.04)%、(48.08±3.77)%]。结论 DDP诱导SKOV3来获得的肿瘤裂解物致敏DC能有效激活抗肿瘤免疫反应,提示化疗与生物治疗联合治疗可能是治疗卵巢癌潜在有效的方法。

Objective To discuss the killing effect of CTL induced by dendritic cells （DCs） sensitized by tumor cell lysate which was acquired by Cisplatin inducing ovarian cancer cells （SKOV 3） to tumor cell. Methods Mononuclear cells of peripheral blood of healthy pepole was separated and the DC was cultivated. There was three groups. Cisplatin group： DC was sensitized by tumor cell lysate which was acquired by Cisplatin inducing ovarian cancer cells. Freeze-thaw groups： DC was sensitized by tumor cell lysate which was obtained by freezing and thawing method. Blank group： There was no any processing. Mature DC and T cells were mixed and cultivated to get cytotoxic T lymphocyte（CTL）. DC phenotype of three groups and CTL subgroup ratio were detected by Flow cytometry （FCM）. The killing effect of CTL to SKOV 3 was tested by MTT. Results The expression of CD 86 and CD 40 of molecular surface marks of DC in Cisplatin group were （76.35±3.99）% and（71.20±6.86）% and it was significantly higher than that of blank group（P〈0.05）, while there was no statistical difference to freeze-thaw group. The ratio of CD 8＋T andCD 4＋T of subgroups of specific CTL induced by two sets of DC were （80.42±4.61）% and （9.55±2.24）%, which was significantly higher than the blank group （P〈0.05）. The kill rate of CTL induced by Cisplatin group to SKOV 3 gradually increased with the increase of E/T ratio （20 ： 1, 40 ： 1, 80 ： 1） and the kill rate [（45.66±3.16）%, （50.16±3.44）%, （58.82±4.06）%] was significantly higher than the freezing and thawing group [（39.71±4.77） %, （47.00±4.02）%, （53.76±4.78）%] and blank group[（37.94±3.74）%,（41.11±5.04）%,（48.08±3.77）%]. Conclusion DC sensitized by tumor cell lysate induced by Cisplatin which was acquired by DDP inducing ovarian cancer cells （SKOV 3） can effectively activate anti-tumor immune response. TIP：The combination therapy of chemotherapy and biological therapy biological combined treatment may be a potential effective method for treatment of ovarian cancer.