骨髓间充质干细胞分泌的外泌体调控肿瘤相关巨噬细胞治疗胰腺癌的实验研究  

Experimental study of exosome derived from bone marrow mesenchymal stem cells in treating pancreatic cancer through regulation of tumor-associated macrophages polarization

作　　者：邵建平[1] 蔡雪军[1] 井树森 苏锋[1] 张立明[1] 李永元 王心[2] Shao Jianping;Cai Xuejun;Jing Shusen;Su Feng;Zhang Liming;Li Yongyuan;Wang Xin(Department of General Surgery,the 5th Central Hospital,Tianjin 300450,China;Department of Surgery,the 4th Central Hospital,Tianjin 300140,China)

机构地区：[1]天津市第五中心医院普通外科,天津300450 [2]天津市第四中心医院外科,天津300140

出　　处：《中华肝胆外科杂志》2022年第12期931-935,共5页Chinese Journal of Hepatobiliary Surgery

基　　金：天津市卫生计生委科技基金面上项目(2014KY04);天津市卫健委科技人才培育项目(RC20093)。

摘　　要：目的探讨骨髓间充质干细胞(BMSC)分泌的外泌体调控肿瘤相关巨噬细胞(TAM)极化对胰腺癌的治疗作用。方法取4周龄C57BL/6雄性小鼠10只, 体重约20 g, 提取BMSC分泌的外泌体并用PKH26标记。取4~6周龄雌性BALB/c-nu/nu品系裸鼠30只, 无特定病原体级, 体重(18.56±0.85)g, 建立胰腺癌裸鼠模型, 并将其分为3组, 每组10只:空白对照组经鼠尾静脉注射磷酸盐缓冲液(PBS);门静脉治疗组经门静脉注射含外泌体的PBS悬液;尾静脉治疗组经尾静脉注射含外泌体的PBS悬液。8周后处死裸鼠, 检测胰腺组织中PKH26表达阳性的细胞百分比, 测定原位肿瘤体积、抑瘤率, 观察肿瘤转移情况、腹水情况、瘤重、瘤肝重。随后检测外周血M1型以及M2型巨噬细胞活化标志物的基因表达、外周血胰腺癌细胞标志物B7-H4、肿瘤糖类抗原199的表达以及胰腺肿瘤组织细胞中Survivin和基质金属蛋白酶-9(MMP-9)的表达。结果门静脉治疗组抑瘤率(72.4±21.6)%与尾静脉治疗组抑瘤率(70.1±20.7)%相比, 差异无统计学意义(t=0.24, P=0.811)。与空白对照组裸鼠相比, 门静脉治疗组与尾静脉治疗组裸鼠的肿瘤体积、瘤重、瘤肝重、肝转移率、肝转移灶数量、腹水发生率和其他器官转移灶总数均降低, 差异具有统计学意义(均P<0.05)。与空白对照组相比, 门静脉治疗组和尾静脉治疗组裸鼠的iNOS、CD68升高, 而Arginase、CD206、B7-H4、肿瘤糖类抗原199、Survivin和MMP-9均降低, 差异具有统计学意义(均P<0.05)。结论 BMSC分泌的外泌体可抑制TAM向M2表型转化, 并诱导其向M1表型转化, 进而抑制胰腺癌的增殖、侵袭、转移能力, 达到抑癌的作用。Objective To investigate the therapeutic effect of exosomes divided from bone marrow mesenchymal stem cell(BMSC)on pancreatic cancer in vivo through regulation of tumor-associated macrophages(TAM)polarization.Methods Ten male C57BL/6 mice weighing approximately 20 g,ages 4 weeks,were used for BMSC exosomes extraction and PKH26 labelling.Thirty female SPF BALB/c-nu/nu nude mice weighing approximately(18.56±0.85)g,ages 4-6 weeks,were adopted for pancreatic carcinoma models.The models were randomly divided into 3 groups with 10 in each:control group(injected with PBS through tail vein),portal vein treatment group(injected with exosomes in PBS through portal vein),tail vein treatment group(injected with exosomes in PBS through tail vein).After the models were executive 8 weeks later,the percentage of PKH26 positive-exosomes in pancreas tissue was quantified by flow cytometry technique.The volume of the primary pancreatic tumor,the tumor volume of inhibitory rate,the number of metastatic nodule,and the ascitic fluid were assess.Also,the weight of liver and the tumor weight were evaluated.The expression of M1 and M2 macrophage-activate biomarkers and the content of pancreatic cancer marker B7-H4 tumor carbohydrate antigen 199 in peripheral blood was detected.Moreover,the expression of Survivin and matrix metalloproteinase-9(MMP-9)in pancreatic cancer tissue cells was also detected.Results There was no significant difference for the tumor volume of inhibitory rate between portal vein treatment group(72.4±21.6)%and tail vein treatment group(70.1±20.7)%(t=0.24,P=0.811).Compared with control group,the volume of the primary pancreatic tumor,the weight of liver,the tumor weight,the number and rate of liver metastatic nodule,the ascitic fluid,and the number of other metastatic nodule were totally lower in the 2 treatment groups with significant difference(all P<0.05).Compared with the control group,iNOS and CD68 in portal vein treatment group and tail vein treatment group were increased,while Arginase,CD206,B7-H4,tumor car

关 键 词：胰腺肿瘤 外泌体 骨髓间充质干细胞 肿瘤相关巨噬细胞 

分 类 号：R735.9[医药卫生—肿瘤]