共培养体系中IL-15通过激活NK细胞ULBP1/NKG2D信号抑制食管癌细胞的迁移和侵袭  

IL-15 inhibits migration and invasion of esophageal cancer cells by activating ULBP1/NKG2D signal of NK cells

作　　者：董良[1] 李洪霖 杨清 段铮[1] 孙明月[1] 许彦超 马纯政(指导)[1] DONG Liang;LI Honglin;YANG Qing;DUAN Zheng;SUN Mingyue;XU Yanchao;MA Chunzheng(Department of Oncology,Henan Provincial Hospital of Traditional Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南省中医院肿瘤科,郑州450000 [2]河南省中医院医务科,郑州450000 [3]河南中医药大学,郑州450000

出　　处：《中国免疫学杂志》2023年第3期466-471,477,共7页Chinese Journal of Immunology

基　　金：国家自然科学基金青年基金项目(81804057);河南省科技厅科技攻关课题(212102311118);河南省中医管理局课题(2019JDZX031);河南省中医药管理局中医临床基地项目(2017JDZX016)。

摘　　要：目的:探讨IL-15对食管癌细胞和自然杀伤(NK)细胞共培养体系中NK细胞活性以及对食管癌细胞迁移和侵袭的影响和机制。方法:qRT-PCR和Western blot检测人正常食管上皮细胞(HEEC)和食管癌细胞系TE-1中IL-15水平。TE-1与NK细胞共培养,共培养体系分为对照组(无处理)、IL-15组(0.2、0.4、0.8μg/ml的IL-15)、IL-15+anti-NKG2D组[UL16结合蛋白/自然杀伤组蛋白2D(ULBP1/NKG2D)的阻断剂anti-NKG2D联合IL-15(0.8μg/ml)]、anti-NKG2D组、IL-15+anti-ASIALO-GM1组[NK细胞抑制剂anti-ASIALO-GM1联合IL-15(0.8μg/ml)]及anti-ASIALO-GM1组。细胞划痕愈合实验和Transwell侵袭实验检测TE-1细胞的迁移和侵袭能力,CCK-8法检测NK细胞增殖率,Western blot检测各组NK细胞表面ULBP1/NKG2D表达,免疫共沉淀技术检测NKG2D与ULBP1的相互作用。结果:TE-1细胞中IL-15水平低于HEEC细胞(P<0.05)。0.2、0.4、0.8μg/ml IL-15处理共培养体系提高NK细胞增殖率,但降低TE-1细胞的迁移率和侵袭率(均P<0.05)。在TE-1与NK细胞的共培养体系中,IL-15组(0.8μg/ml)NK细胞膜蛋白ULBP1和NKG2D水平较对照组上调(均P<0.05);anti-NKG2D抑制了NKG2D表达,并抑制了NKG2D与ULBP1的相互作用,与IL-15(0.8μg/ml)组相比,IL-15+anti-NKG2D组NKG2D表达水平和NK细胞增殖率均降低(均P<0.05),但TE-1细胞的迁移率和侵袭率升高(均P<0.05),且NK细胞抑制剂anti-ASIALO-GM1与anti-NKG2D的作用效果一致。结论:IL-15通过激活细胞表面ULBP1/NKG2D信号促进NK细胞活性,从而抑制食管癌细胞的迁移和侵袭。Objective:To investigate effect of IL-15 on activity of natural killer(NK)cells and migration and invasion of esophageal cancer cells in co-culture system of esophageal cancer cells and NK cells and its mechanism.Methods:Levels of IL-15 in human normal esophageal epithelial cells(HEEC)and esophageal carcinoma cells TE-1 were detected by qRT-PCR and Western blot.TE-1 and NK cells were co-cultured,the co-culture system was divided into control group(no treatment),IL-15 group(0.2,0.4,0.8μg/ml IL-15),IL-15+anti-NKG2D group[UL16 binding protein 1/natural killer group 2 member D(ULBP1/NKG2D)blocler anti-NKG2D combined with IL-15(0.8μg/ml)],anti-NKG2D group,IL-15+anti-ASIALO-GM1 group[anti-ASIALO-GM1 combined with IL-15(0.8μg/ml),an inhibitor of NK cells]and anti-ASIALO-GM1 group.Migration and invasion ability of TE-1 cells were detected by scratch healing assay and Transwell invasion assay.Proliferation rate of NK cells was detected by CCK-8 assay.Expression of ULBP1/NKG2D on surface of NK cells in each group was detected by Western blot,co-immunoprecipitation technique was used to detect interaction between NKG2D and ULBP1.Results:Level of IL-15 in TE-1 cells was lower than that in HEEC cells(P<0.05).0.2,0.4,0.8μg/ml IL-15 co-culture system increased proliferation rate of NK cells,but decreased migration rate and invasion rate of TE-1 cells(all P<0.05).In co-culture system of TE-1 and NK cells,levels of NK cell membrane proteins ULBP1 and NKG2D in IL-15(0.8μg/ml)group were up-regulated compared with control group(all P<0.05).Anti-NKG2D inhibited expression of NKG2D and blocked interaction between NKG2D and ULBP1.Compared with IL-15(0.8μg/ml)group,expression level of NKG2D and proliferation rate of NK cells were decreased in IL-15+anti-NKG2D group(all P<0.05).However,migration rate and invasion rate of TE-1 cells were increased(both P<0.05),and effect of anti-ASIALO-GM1,an inhibitor of NK cells,was consistent with that of anti-NKG2D.Conclusion:IL-15 inhibits migration and invasion of esophageal cancer cells by ac

关 键 词：白介素-15 食管癌细胞 迁移 侵袭 自然杀伤细胞 自然杀伤组2成员D蛋白/配体1 

分 类 号：R735.1[医药卫生—肿瘤]