异基因骨髓细胞联合淋巴细胞输注抑制结肠癌肝转移  

Inhibiting colorectal cancer liver metastasis by combined injection of allogeneic bone marrow cells and lymphocytes

作　　者：瞿元乾 周庆[1] 张进[1] 徐三荣[1] Qu Yuan-qian;Zhou Qing;Zhang Jin;Xu San-rong(Department of General Surgery,Affiliated Hospital of Jiangsu University,Zhenjiang 212000,Jiangsu Province,China)

机构地区：[1]江苏大学附属医院普外科,江苏省镇江市212000

出　　处：《中国组织工程研究》2018年第25期4053-4058,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81301693);江苏省自然科学基金项目(BK20130474);镇江市社会发展项目(SH2015056)~~

摘　　要：背景:异基因造血干细胞移植对结肠癌术后肝转移的预防和治疗在临床以及动物实验中鲜有研究报道,结肠癌术后肿瘤负荷较小,此时建立混合型嵌合体可望取得一定疗效。目的:探讨异基因造血干细胞移植对结肠癌肝转移的抑制作用及其对嵌合体和移植物抗宿主病的影响,并对其抑瘤机制进行研究。方法:BALB/c×C57BL/6杂交一代CB6F1小鼠保脾注射CT-26大肠癌细胞建立结肠癌肝转移模型,随机分为4组:对照组、环磷酰胺组、全相合脾细胞加骨髓细胞移植+环磷酰胺组(全相合移植组)、半相合脾细胞加骨髓细胞移植+环磷酰胺组(半相合移植组),每组8只。取自C57BL/6小鼠和CB6F1小鼠骨髓及脾细胞混合后行尾静脉注射,之后接受腹腔注射环磷酰胺进行非清髓性预处理,再输注供鼠淋巴细胞。观察各组小鼠生存时间、肿瘤肝转移以及移植物抗宿主病发生情况;流式细胞仪进行嵌合体分析,ELISA方法测定血浆白细胞介素2、γ-干扰素、白细胞介素4、转化生长因子β水平。结果与结论:(1)半相合移植组小鼠生存期明显延长,结肠癌肝转移抑制率明显增高,抑瘤率明显增加,与环磷酰胺组及全相合移植组相比,差异有显著性意义;(2)半相合移植组小鼠混合细胞移植7 d后,供鼠细胞嵌合率逐渐增加;至移植后28 d,供鼠细胞基本取代受鼠细胞,嵌合率达99%以上;(3)半相合移植组小鼠的移植物抗肿瘤效应明显增强,但未出现严重的移植物抗宿主病症状,且小鼠血浆白细胞介素2、γ-干扰素水平较其他组明显升高,转化生长因子β水平明显降低,而白细胞介素4水平无明显差异;(4)结果表明,小鼠经环磷酰胺非清髓处理的异基因骨髓细胞联合淋巴细胞移植后嵌合率明显增加,伴随产生了明显的移植物抗肿瘤效应,其对结肠癌肝转移具有抑制作用。血浆白细胞介素2、γ-干扰素、转化生长因子β水平变化与肿瘤�BACKGROUND:Clinical and experimental studies on prevention and treatment of hepatic metastasis after colonic cancer treatment by allogeneic hematopoietic stem cell transplantation have yet to be well reported.A mixed chimera is expected to achieve certain effects in patients with low-tumor burden after colorectal cancer resection.OBJECTIVE:To investigate the inhibitory effect of allogeneic hematopoietic stem cell transplantation on colorectal cancer liver metastasis as well as the effect on chimera level and graft-versus-host disease and to study the anti-tumor mechanism.METHODS:CB6F1 mice were hybridized by BALB/c×C57BL/6 mice and injected with CT-26 cells to make animal models of colorectal cancer liver metastasis.Then,model mice were randomized into four groups(n=8 per group):control group,cyclophosphamide group,HLA-identical spleen cells plus bone marrow cell transplantation+cyclophosphamide group(HLA-identical transplantation group),and haploidentical spleen cells plus bone marrow cell transplantation+cyclophosphamide group(haploidentical transplantation group).Mixture of spleen cells and bone marrow cells from CB6F1 and C57BL/6 mice were injected via the tail vein followed by a non-myeloablative pretreatment with intraperitoneal injection of cyclophosphamide,and thereafter,lymphocytes were given via infusion.Survival time,tumor liver metastasis and the occurrence of graft-versus-host disease were observed in mice.The chimera was analyzed by flow cytometry,and levels of plasma interleukin-2,interferon-γ,interleukin-4,and transforming growth factorβwere determined by ELISA RESULTS AND CONCLUSION:The survival time and inhibition rate of liver metastasis in the haploidentical transplantation group were significantly prolonged and increased compared with cyclophosphamid and HLA-identical transplantation group,respectively.At 7 days after transplantation,the chimera level in the haploidentical transplantation group was significantly increased up to over 99%at 28 days of transplantation,indicating that donor ce

关 键 词：结肠癌肝转移 异基因 骨髓细胞 脾细胞 淋巴细胞 移植物抗宿主病 嵌合体 小鼠 干细胞 国家自然科学基金 结肠肿瘤 肿瘤转移 肝 骨髓移植 淋巴细胞 嵌合体 移植物抗宿主病 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]