肺癌细胞致敏树突状细胞疫苗的体内外抗肿瘤作用  

In vitro and in vivo anti-tumor effects of dendritic cell vaccines sensitized by NCI-H460 lung cancer cell line

作　　者：傅永强[1] 张在云[2] 柳月安[3] 李旭升[4] 

机构地区：[1]金华职业技术学院医学院内科,浙江金华321017 [2]山东大学附属第二医院肿瘤科,济南250033 [3]山东省肿瘤医院药剂科,济南250117 [4]金华职业技术医学院基础医学部,浙江金华321017

出　　处：《中国肿瘤生物治疗杂志》2007年第6期562-566,共5页Chinese Journal of Cancer Biotherapy

基　　金：金华市科技创新重点项目(No.2004-2-302)~~

摘　　要：目的:研究以NCI-H460肺癌细胞致敏的树突状细胞(Dendritic cells,DCs)疫苗的抗肿瘤作用。方法:将IL-18基因转染的NCI-H460细胞及未转染NCI-H460细胞与DC融合作为转染融合DC组及融合DC组疫苗,以NCI-H460细胞RNA冲击的DC为冲击DC组疫苗,以未冲击的DC为DC组疫苗。以MTT法检测4组DC疫苗刺激T细胞增殖的作用,用ELISA法测定DC疫苗上清液中IL-12的含量,用LDH法测定DC疫苗对NCI-H460细胞的杀伤作用。裸鼠皮下接种NCI-H460细胞及DC疫苗,观察成瘤时间及裸鼠存活情况;荷瘤裸鼠瘤内注射DC疫苗,比较肿瘤体积。结果:4组DC疫苗均能刺激T细胞增殖,刺激作用强度为转染融合DC组>融合DC组>冲击DC组>DC组;4组均能分泌IL-12,转染融合DC组IL-12的分泌量高于融合DC组,冲击DC组高于DC组;转染融合、融合、冲击DC和DC组疫苗对NCI-H460细胞的杀伤率分别为79.73%、50.68%、35.81%及4.05%。转染融合组裸鼠移植瘤成瘤时间[(12.82±2.85)d]长于冲击DC组[(8.52±1.97)d](P<0.05)和DC组[(8.33±1.63)d](P<0.01);移植瘤内注射疫苗后的肿瘤体积比较,转染融合组<融合组<冲击DC组<DC组。结论:以NCI-H460肺癌细胞致敏的DC疫苗在体外及裸鼠体内均显示了抗肿瘤作用,转染融合DC疫苗抗肿瘤作用最强,融合和RNA冲击DC疫苗次之,单纯DC疫苗作用最弱。Objective： To study the anti-tumor effects of dendritic cell（DC） vaccines sensitized by NCI-H460 lung cancer cell lines. Methods： The study consisted of 4 groups： IL-18/fusion group, fusion group, pulsed DC group and DC group. IL-18 gene transfected or non-transfected NCI-H460 cells were fused with DC and the fused cells served as DC vac- cines for IL-18/fusion group and fusion group, respectively. DC pulsed with RNA of NCI-H460 cells were used for pulsed DC group and non-pulsed DC for DC group. T cell proliferation after stimulation with 4 vaccines was tested. IL-12 levels in the supernatants of DC vaccines were measured with ELISA. LDH method was used to evaluate the in vitro anti-tumor effect of DC vaccines. NCI-H460 cancer cells and DC vaccines were subcutaneously injected to nude mice; the tumor growth and survival of nude mice were observed. Tumor-loaded nude mice were inoculated with DC vaccines and tumor sizes of the 4 groups were compared. Results： The T cell proliferation activity of the 4 groups was： IL-18/fusion group 〉 fusion group 〉 pulsed DC group 〉 DC group. IL-12 secretion in IL-18/fusion group was higher than that in the fusion group, and IL-12 in the pulsed DC group was higher than that in the DC group. The in vitro killing rates of the 4 groups were 79. 73% ,50. 68% ,35.81% and 4.05% , respectively. Tumor forming time in IL-18/fusion group（ [ 12.82 ±2.85 ] d） was longer than those in the pulsed DC group （ [ 8.52 ± 1.97 ] d, P 〈 0. 05 ） and DC group （ [ 8.33 ± 1.63 ] d, P 〈 0. 01 ）.The tumor sizes of the 4 groups were： IL-18/fusion group 〈 fusion group 〈 pulsed DC group 〈 DC group. Conclusion： DC vaccines sensitized by NCI-H460 lung cancer cell line have anti-tumor effects. Among the 4 vaccines, IL-18/fusion group have the most powerful anti-tumor activity, followed by fusion group. The anti-tumor activity of DC group is the weakest one.

关 键 词：树突状细胞 疫苗 肺癌细胞 细胞融合 白细胞介素-18 

分 类 号：R734.2[医药卫生—肿瘤] R730.54[医药卫生—临床医学]