脐血来源NK细胞对肝肿瘤生长抑制作用  

Inhibitory effect of NK cells derived from cord blood on liver tumor growth

作　　者：吴炳义 李娟华[4] 李烨 张兰兰 朱伟[5] 陈琼[2,3] 吕成伟[2,3] WU Bingyi;LI Juanhua;LI Ye;ZHANG Lanlan;ZHU Wei;CEHN Qiong;LV Chengwei(Xiangxue Southern Precision Medicine Technology Co.,Guangzhou,Guangdong,China,510063;Medical Research Center,Nanfang Hospital,Southern Medical University,Guangzhou,Guangdong,China,510515;Precision Medicine Center,Nanfang Hospital,Southern Medical University,Guangzhou,Guangdong,China,510515;Obstetrics and Gynecology Department,Zengcheng Branch of Nanfang Hospital,Guangzhou,Guangdong,China,510000;Liver Disease Medical Center,Nanfang Hospital,Southerm Medical University,Guangzhou,Guangdong,China,510515)

机构地区：[1]广州香雪南方精准医学科技有限公司,广东广州510663 [2]南方医科大学南方医院医学实验中心,广东广州510515 [3]南方医科大学南方医院精准医学中心,广东广州510515 [4]南方医院增城分院妇产科,广东广州510000 [5]南方医科大学南方医院肝病中心,广东广州510515

出　　处：《分子诊断与治疗杂志》2022年第9期1484-1488,共5页Journal of Molecular Diagnostics and Therapy

基　　金：国家自然科学基金(81771340);横向课题(HXKY201809070203)。

摘　　要：目的 培养脐血来源NK细胞,通过细胞和动物水平验证其用于肝癌治疗的安全性和有效性。方法 NK细胞体外诱导培养,观察培养过程细胞形态、细胞总数、增殖倍数,流式检测其中CD3~-CD56~+细胞的比例,细胞毒性检测验证NK细胞对癌细胞的杀伤活性;建立肝癌裸鼠皮下荷瘤模型,采用瘤内与静脉方式注射NK细胞,密切观察不同时间段实验组和对照组裸鼠移植瘤大小、体积、质量以及裸鼠存活状态。荷瘤40天后取肿瘤组织进行分析。通过流式分析、HE染色和免疫荧光检测NK细胞移植在裸鼠体内第37天含量,分析NK细胞在裸鼠体内残留情况。结果 脐血分离单个核细胞经体外诱导培养的NK细胞能保持典型的NK细胞表型特征,细胞活性好、增殖能力强、纯度高,对多种癌细胞系均具有良好的杀伤作用。与对照组相比,NK细胞注射组均能抑制肝肿瘤生长。NK细胞移植35天后,尾静脉组肿瘤大小显著下降(P<0.05)。同时NK细胞在体内无明显残留,表明NK细胞小鼠体内移植无致瘤性。结论 本研究制备的脐血NK细胞符合临床研究需求,可作为肝癌患者临床治疗的替代方案,为下一步开展临床研究提供前期基础。Objective To culture the NK cells derived from cord blood and verify the safety and efficacy of NK cells in the treatment of liver cancer at cellular and animal levels. Methods NK cells were induced and cultured in vitro,and the cell morphology,total number of cells,and multiplication factor were observed during the culture process,and the proportion of CD3CD56cells was detected by flow cytometry. The cytotoxicity assay was performed to verify the killing effect of NK cells on various cancer cells in vitro. A subcutaneous tumor-bearing model of liver cancer nude mice was established,and NK cells were injected intratumorally and intravenously. The size,volume,mass and survival status of the transplanted tumors in the experimental group and the control group were observed in different time periods. Tumor tissue was taken for K analysis 40 days after tumor bearing. The content of NK cells transplanted in nude mice on the 37th day was detected by flow analysis,HE staining and immunofluorescence,and the residual NK cells in nude mice were analyzed. Results The NK cells induced and cultured by umbilical cord blood isolated mononuclear cells can maintain the typical NK cell phenotype characteristics,with good cell activity,strong proliferation ability and high purity,and have a good killing effect on a variety of cancer cell lines. Compared with the control group,the NK cell injection group could inhibit the growth of liver tumor. After 35 days of NK cell transplantation,the tumor size in the tail vein injected group decreased significantly(P<0.05). At the same time,there were no obvious residual NK cells in the body,which indicated that the transplantation of NK cells into mice had no tumorigenicity. Conclusion The cord blood NK cells prepared in this study meet the needs of clinical research and can be used as an alternative for clinical treatment of liver cancer patients,providing a preliminary basis for the next clinical research.

关 键 词：自然杀伤细胞 肝癌 脐血 动物试验 

分 类 号：R735.7[医药卫生—肿瘤]