射频消融与细胞因子诱导的杀伤细胞协同抗肿瘤效应的研究  

The synergistic anti -tumor effect of radiofrequency and cytokine -induced killer cells in mice

作　　者：戴夕超[1] 石亮荣[1] 孙明芬 徐斌[2] 李晓东[1] 陈陆俊[2] 胡文蔚[1] 朱一蓓[3] 蒋敬庭[2] 吴昌平[1] 

机构地区：[1]苏州大学附属第三医院肿瘤科,常州213003 [2]苏州大学附属第三医院肿瘤生物诊疗中心,常州213003 [3]苏州大学生物技术研究所,215006

出　　处：《中华实验外科杂志》2016年第6期1552-1556,共5页Chinese Journal of Experimental Surgery

基　　金：基金项目：国家自然科学基金（81201741、31570877、31570908、81301960、81171653、31428005）;江苏省重点研发计划与社会发展项目（BE2015633、BE2015634）

摘　　要：目的观察射频消融（RFA）联合细胞因子诱导的杀伤细胞（CIK）的抗肿瘤免疫效应。方法建立小鼠双侧背部B16黑色素瘤与CT26结肠癌皮下移植瘤模型，成瘤后分d组，分别予以单独右侧肿瘤RFA、CIK治疗、两者联合治疗或未治疗对照，记录左侧肿瘤大小与小鼠生存时间。在B16移植瘤模型中分析各治疗组对侧肿瘤内淋巴细胞亚群比例与数量，并检测浸润至对侧肿瘤内的CIK细胞的数量与表型。结果RFA与CIK单独治疗均可产生短暂的肿瘤抑制作用，而联合治疗能显著抑制对侧肿瘤生长；联合治疗组中位生存期为（42．0±3．3）d，显著长于未治疗对照组[（20．5±2．2）d]、RFA组[（26．0±2．2）d]与CIK单独治疗组[（31．5±2．2）d，P〈0．01]；联合治疗组小鼠对侧肿瘤内浸润淋巴细胞总数增加，其中每克肿瘤组织内CD8^＋T细胞、CD4^＋T细胞、自然杀伤（NK：CD3^-NK1．1^＋）细胞和自然杀伤T细胞（NKT：CD5^＋NK1．1^＋）数量均显著增加，分别为（2．41±1．03）×10^6cell／g、（4．52±1．63）×10^cell／g、（1．62±0．31）×10^＋cell／g、（0．42±0．08）×10^＋cell／g；CD8^＋／调节性T细胞[Treg：CD4^＋叉状头／翅膀状螺旋转录因子（Foxp3）^＋]比值增大为（3．85±0．63）。髓源性抑制细胞（MDSC）比例降低，为（8．92±3．05）％。RFA联合CIK组较外科切除联合CIK组小鼠对侧肿瘤内外源性CIK细胞聚集增加，而细胞核增殖抗原（Ki-67）、可诱导共刺激分子（ICOS）与颗粒酶B（GranzymeB）的表达水平上调，分别为（79．12±1．51）％、（56．13±2．12）％、（63．51±1．82）％。结论RFA治疗可促进外源性的CIK细胞向消融区外肿瘤内迁移，并提高CIK细胞在肿瘤内增殖能力与活性，RFA联合CIK治疗可增强消融区外肿瘤微环境中抗肿瘤免疫效应，产生协同抗肿瘤作用。Objective This study aimed to evaluate the anti - tumor immunity of the combination treatment with radiofrequency ablation （RFA） and cytokine - induced killer cells （CIK）. Methods Mice were inoculated with B16 melonoma or CT26 colon cancer cell line on bilateral flanks, and randomly divid- ed into four groups in which RFA for tumor right flank, CIK cell infusion, RFA plus CIK and no interven- tion was administrated, respectively. The growth curve of the left tumor and survival curve of mice were re- corded. The amount and subset of the lymphocytes infiltrating into the tumor on the left flank were analyzed by flow cytometry in B16 tumor - bearing mice. In addition, the number and phenotype of adoptive CIK mi- grating into the tumor were tested. Results A short - lived inhibition of tumor growth was observed in mice treated with RFA or CIK alone. Combination treatment with RFA and CIK significantly inhibited the growth of tumors on contralateral flank. The median survival of mice treated with RFA and CIK was （42. 0 ± 3.3 ） d, which was significant longer than that in untreated mice （20. 5 ± 2. 2 ） d, RFA alone （26. 0 ± 2. 2） d, CIK alone （31.5 ± 2. 2） d （ P 〈 0. 01 ）. The number of immune ceils, including CD8 ^＋ , CD4 ^＋ T, natural killer （ NK： CD3 ^- NK1.1^＋ ） and NKT （ CD3^＋ NK1.1^＋ ）, infiltrating into the contralateral tumor, was respectively （2. 41 ± 1.03） × 10^6 cell/g tumor, （4. 52 ± 1.63） × 10^6 cell/g tumor, （ 1.62 ± 0. 31） × 10^6 cell/g tumor, （0. 42 ±20. 08） × 10^6 cell/g tumor, significantly increased in mice treated with RFA plus CIK, as comparing to mice treated with RFA and CIK alone. An elevated CD8^＋/regulatory T cells [ Treg： CD4^＋ forkhead box P3 （Foxp3）^＋ ] ratio （3.85 ± 0. 63 ） and decreased accumulation of mye- loid - derived suppressor cells （ MDSC）, which the frequency in tumor was （ 8. 92 ± 3.05 ） %, were ob- served in RFA plus CIK group. Compared to mice treated with surgery plus CIK, t

关 键 词：射频消融 细胞因子诱导的杀伤细胞 结肠癌 黑色素瘤 抗肿瘤免疫效应 

分 类 号：R730.5[医药卫生—肿瘤]