白血病细胞外泌体miR-17-5p抑制NK细胞杀伤敏感性并促进白血病细胞免疫逃逸  

Leukemia cell-derived exosomal miR-17-5p inhibits NK cell cytotoxic sensitivity and promotes immune evasion of leukemia cell

作　　者：罗靖 罗政权 谷晓辉 王云霞 李娜 Luo Jing;Luo Zhiquan;Gu Xiaohui;Wang Yunxi;Li Na(Department of Clinical Laboratory/Department of Pathology,Mingren Hospital,Handan 056000,China;Department of Pathology,Fengfeng General Hospital,North China Medical and Health Group,Handan 056200,China;Zhengzhou Jinyu Clinical Laboratory Center,Zhengzhou 450000,China)

机构地区：[1]邯郸明仁医院检验科/病理科,邯郸056000 [2]华北医疗健康集团峰峰总医院病理科,邯郸056200 [3]郑州金域临床检验中心,郑州450000

出　　处：《中国组织化学与细胞化学杂志》2024年第6期540-546,共7页Chinese Journal of Histochemistry and Cytochemistry

摘　　要：目的探讨白血病细胞外泌体(exosome,Exos)miR-17-5p对NK细胞杀伤敏感性的调控作用及其对免疫逃逸的影响。方法分别提取人正常骨髓细胞HS-5、白血病细胞Jurkat和HL-60以及转染miR-17-5p-si RNA-NC或miR-17-5p-si RNA的Jurkat细胞的Exos。采用试剂盒测定NK-92MI细胞对白血病细胞的杀伤毒性;定量荧光PCR(q RT-PCR)检测Exos中miR-17-5p水平;双荧光素酶报告实验验证miR-17-5p与MHCI类链相关蛋白A和B(MICA/B)的结合;Westernblot检测MICA/B蛋白表达。结果Jurkat、HL-60细胞提取的Exos与NK-92MI细胞共孵育后,Jurkat、HL-60细胞LDH含量、细胞凋亡率均降低,miR-17-5p水平增高。转染miR-17-5p-si RNA的Jurkat细胞提取的Exos干预Jurkat细胞后,Jurkat细胞中LDH含量、细胞凋亡率、MICA/B蛋白水平增高。MICA/B与miR-17-5p可靶向结合。结论敲低白血病细胞Exos的miR-17-5p可上调MICA/B表达,增加白血病细胞对NK细胞杀伤的敏感性,抑制白血病细胞免疫逃逸。Objective To explore the regulatory effect of leukemia cell-derived exosome(Exos)miR-17-5p on NK cell cytotoxicity and its impact on immune evasion.Methods Exosomes were extracted from human normal bone marrow cell line HS-5,leukemia cell lines Jurkat and HL-60,and Jurkat cells transfected with miR-17-5p-siRNA-NC or miR-17-5p-siRNA.NK-92MI cell cytotoxicity against leukemia cells was assessed using a cytotoxicity assay kit.The level of miR-17-5p in Exos was quantified by qRT-PCR.A dual-luciferase reporter assay was performed to validate the interaction between miR-17-5p and MHC class I chain-related proteins A and B(MICA/B).Western blot analysis was used to detect MICA/B protein expression.Results After co-culturing NK-92MI cells with Exos from Jurkat and HL-60 cells,a decrease in LDH levels and apoptosis rates in Jurkat and HL-60 cells was observed,accompanied by an increase in miR-17-5p levels.Exosomes extracted from Jurkat cells transfected with miR-17-5p-siRNA increased LDH levels,apoptosis rates,and MICA/B protein expression in Jurkat cells.MICA/B was found to be a target of miR-17-5p binding.Conclusion Knockdown of miR-17-5p in leukemia cell-derived Exos can upregulate MICA/B expression,enhance leukemia cell sensitivity to NK cell cytotoxicity,and suppress immune evasion of leukemia cells.

关 键 词：白血病 外泌体 miR-17-5p NK细胞 MHCⅠ类链相关蛋白A/B 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学]