NK细胞特异性敲除UTX以性别差异方式调控黑色素瘤肺转移  

NK cell-specific knockout of UTX modulates pulmonary metastasis of melanoma in a sex-dependent manner

作　　者：黄沛 王洪晨 黄河 谢佳新 吴玉[1] 周思敏[1] 廖馨怡 管潇 HUANG Pei;WANG Hongchen;HUANG He;XIE Jiaxin;WU Yu;ZHOU Simin;LIAO Xinyi;GUAN Xiao(Department of High Altitude Operational Medicine,Faculty of High Altitude Military Medicine,Army Medical University(Third Military Medical University),Chongqing;Department of Clinical Hematology,Faculty of Pharmacy and Laboratory Medicine,Army Medical University(Third Military Medical University),Chongqing;Department of Neurology,No.940 Hospital of Joint Logistics Support Force,Lanzhou,Gansu;Chongqing Medical and Pharmaceutical College,Chongqing,China)

机构地区：[1]陆军军医大学(第三军医大学)高原军事医学系高原作业医学教研室,重庆 [2]陆军军医大学(第三军医大学)药学与检验医学系临床血液学教研室,重庆 [3]联勤保障部队第九四〇医院神经内科,甘肃兰州 [4]重庆医药高等专科学校,重庆

出　　处：《陆军军医大学学报》2025年第8期807-815,共9页Journal of Army Medical University

基　　金：国家自然科学基金青年项目(82204398);重庆市自然科学基金面上项目(CSTB2022NSCQ-MSX0410);联勤保障部队第九四〇医院高层次培养人才工程年轻储备力量项目(2024-G3-6)。

摘　　要：目的探究X染色体编码的表观调节因子UTX对自然杀伤(natural killer,NK)细胞抗肿瘤活性的调控作用。方法采用Ncr1-i Cre工具雄鼠与UTX^(fl/fl)雌鼠杂交,获得F1代Ncr1-i Cre^(+)UTX^(fl/-)雄鼠,进而将F1代小鼠与UTX^(fl/fl)雌鼠杂交,获得对照雄鼠Ncr1-i Cre^(-)UTX^(fl/-)(M-对照)和NK细胞特异性敲除UTX雄鼠Ncr1-i Cre^(+)UTX^(fl/-)(M-敲除);以及对照雌鼠Ncr1-i Cre^(-)UTX^(fl/fl)(F-对照)和UTX基因敲除雌鼠Ncr1-i Cre^(+)UTX^(fl/fl)(F-敲除)。UTX基因敲除小鼠通过尾静脉注射黑色素瘤细胞B16F10,观察肺脏转移的肿瘤结节数量;并通过流式细胞术分析肺脏NK细胞比例(CD3~-CD19~-NK1.1^(+))、数量、成熟分子KLRG1和CD11b、表达活化受体NKG2D和CD69及效应分子穿孔素、颗粒酶B、CD107a和干扰素-γ(interferon-γ,IFN-γ)的改变;继而将分选的NK细胞与B16F10细胞共培养,通过流式细胞术检测B16F10细胞凋亡的改变。结果与M-对照雄鼠相比较,M-敲除雄鼠肺脏黑色素瘤结节数量显著减少(P<0.05);肺脏NK细胞比例和数量均显著增加(P<0.01);NK细胞成熟标志分子KLRG1和CD11b比例无显著改变;NK细胞毒性分子穿孔素表达增加(P<0.01),而效应分子颗粒酶B、脱颗粒标志分子CD107a,以及细胞因子IFN-γ无统计学差异;NK细胞与B16F10细胞共培养导致肿瘤细胞凋亡显著增加(P<0.05)。相比于F-对照雌鼠,F-敲除雌鼠肺脏黑色素瘤结节数量无统计学差异;肺脏NK细胞比例和数量显著增加(P<0.05);KLRG1和CD11b比例显著减少(P<0.01);NK细胞穿孔素表达增加,颗粒酶B、CD107a和IFN-γ表达均显著减少(P<0.01);NK细胞与B16F10细胞共培养,肿瘤细胞凋亡显著减少(P<0.05)。结论NK特异性敲除UTX以性别差异方式调控小鼠黑色素瘤肺转移。Objective To explore the role of X chromosome encoded epigenetic regulator UTX in NK cell-mediated anti-tumor activity.Methods Male Ncr1-iCre mice were crossed with female UTX^(fl/fl) mice to generate F1 Ncr1-iCre^(+)UTX^(fl/-)male mice,which were further crossed with female UTX^(fl/fl) mice to obtain male Ncr1-iCre^(-)UTX ^(fl/-)control mice(M-Con)and NK-specific deletion of UTX male mice Ncr1-iCre^(+)UTX^(fl/-)(M-KO),as well as female Ncr1-iCre^(-)UTX^(fl/fl) control mice(F-Con)and UTX-deficient female mice Ncr1-iCre^(+)UTX^(fl/fl)(F-KO).UTX-deficient mice were injected with melanoma cell line B16F10 via tail vein to observe pulmonary metastatic tumor nodules.Moreover,flow cytometry was applied to detect the proportion and quantity of pulmonary NK cells(CD3-CD19-NK1.1^(+)),maturation makers KLRG1 and CD11b,activation receptors NKG2D and CD69,and effector molecules,including perforin,granzyme B,CD107a,and IFN-γ.Then pulmonary NK cells were sorted and co-cultured with B16F10 cells,and the apoptosis of the melanoma cells was measured with flow cytometry.Results Compared with the M-Con mice,the M-KO mice presented less number of pulmonary tumor nodules(P<0.05),increased proportion and quantity of NK cells in the tumor microenvironment(P<0.01),though no obvious changes in the ratio of NK maturation makers KLRG1 to CD11b,enhanced expression level of cytotoxic molecule perforin(P<0.01),but no changes in the expression of effector molecule granzyme B,degranulation marker CD107a and cytokine IFN-γin NK cells.Co-culture of NK cells and B16F10 cells promoted the apoptosis of tumor cells(P<0.05).Compared with the F-Con mice,the F-KO mice had no statistical difference in the number of pulmonary tumor nodules,but larger proportion and number of NK cells(P<0.05),decreased ratio of KLRG1 to CD11b(P<0.01),elevated level of perforin but decreased levels of granzyme B,CD107a and IFN-γin NK cells(P<0.01).The co-culture of NK cells and B16F10 cells reduced the apoptosis of tumor cells in F-KO female mice(P<0.05).Conclusion NK-sp

关 键 词：肺转移 黑色素瘤 NK细胞 性别 UTX 

分 类 号：R73-362[医药卫生—肿瘤] R734.2[医药卫生—临床医学] R739.5