根皮素对NK细胞杀伤人胃癌SGC-7901细胞的增强效应  

Enhanced effect of phloretin on killing effect of NK cells on human gastric cancer SGC-7901 cells

作　　者：耿天祥[1] 王娅菲[1] 张杰 GENG Tianxiang;WANG Yafei;ZHANG Jie(Department of Gastrointestinal Surgery,Jiaozuo People's Hospital,Jiaozuo,Henan 454000,China)

机构地区：[1]焦作市人民医院胃肠外科,河南焦作454000

出　　处：《热带医学杂志》2023年第4期470-475,I0001,共7页Journal of Tropical Medicine

基　　金：河南省医学科技项目科研课题(LHGJ20190371)。

摘　　要：目的探究根皮素对自然杀伤(NK)细胞杀伤人胃癌(GC)SGC-7901细胞的增强效应。方法采用0、4、8和16μmol/L的根皮素处理NK细胞和人胃癌SGC-7901细胞,MTT法检测NK细胞、SGC-7901细胞增殖情况;流式细胞术检测SGC-7901细胞NKG2D配体(MICA、MICB、ULBP1、ULBP2)表达情况;免疫荧光检测SGC-7901细胞DNA损伤程度。将SGC-7901细胞分为对照组(NC组)、根皮素组(8μmol/L)、NK细胞组(10∶1)、根皮素+NK细胞组(8μmol/L根皮素+10∶1 NK细胞),钙黄绿素染色法检测NK细胞对SGC-7901细胞的杀伤率;ELISA试剂盒检测培养液中肿瘤坏死因子-α(TNF-α)和人干扰素-γ(IFN-γ)含量。每组数据设置3个技术重复,6个生物学重复,结果为6个生物学重复的均值。结果与根皮素(0μmol/L)组[24 h:(100.59±5.95)%,48 h:(100.59±4.75)%]比较,根皮素(4μmol/L)组[24 h:(84.78±7.45)%,48 h:(74.59±5.43)%]、根皮素(8μmol/L)组[24 h:(59.34±6.04)%,48 h:(50.34±4.62)%]、根皮素(16μmol/L)组[24 h:(42.32±5.12)%,48 h:(28.32±2.37)%]SGC-7901细胞的增殖率依次降低,差异均有统计学意义(F=89.771、214.322,P均<0.05)。与根皮素(0μmol/L)组比较,根皮素(4μmol/L)组、根皮素(8μmol/L)组、根皮素(16μmol/L)组处理24、48 h后,NK细胞增殖率均显著升高,MICA、MICB、ULBP1、ULBP2、γ-H2AX水平依次升高,差异均有统计学意义(P均<0.05)。与NC组比较,根皮素组、NK组、NK+根皮素组杀伤率,培养液中TNF-α和IFN-γ含量依次增高(F=330.181、288.215、336.648,P均<0.05)。结论根皮素可增强NK细胞对GC细胞的杀伤活性,可能与破坏GC细胞DNA,升高细胞膜NKG2D配体水平,促进TNF-α和IFN-γ释放有关。Objective To explore the enhancing effect of phloretin on natural killer(NK)cells killing human gastric cancer SGC-7901 cells.Methods NK cells and human gastric cancer SGC-7901 cells were treated with 0,4,8 and 16μmol/L phloretin and the proliferation of NK cells and SGC-7901 cells was detected by MTT method;the expression of NKG2D ligands(MICA,MICB,ULBP1,ULBP2)in SGC-7901 cells was detected by flow cytometry;the degree of DNA damage in SGC-7901 cells was detected by immunofluorescence.The SGC-7901 cells were grouped into control group(NC group),phloretin group(8μmol/L),NK cell group(10∶1),and phloretin+NK cell group(8μmol/L phloretin+10∶1 NK cells);calcein staining was used to detect the killing rate of NK cells to SGC-7901 cells;ELISA kits were used to detect the contents of tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ)in the culture medium.Each group of data was set with 3 technical replicates and 6 biological replicates,and the result was the average of 6 biological replicates.Results Compared with the phloretin(0μmol/L)group[24 h:(100.59±5.95)%,48 h:(100.59±4.75)%],the proliferation rate of SGC-7901 cells decreased sequentially in the phloretin(4μmol/L)group[24 h:(84.78±7.45)%,48 h:(74.59±5.43)%],the phloretin(8μmol/L)group[24 h:(59.34±6.04)%,48 h:(50.34±4.62)%],and the phloretin(16μmol/L)group[24h:(42.32±5.12)%,48 h:(28.32±2.37)%(F=89.771,214.322,all P<0.05)],and the proliferation rate of NK cells increased obviously(F=26.707,23.295,all P<0.05);the levels of MICA,MICB,ULBP1,ULBP2,andγ-H2AX increased sequentially(F=275.107,270.382,156.905,143.715,271.311,all P<0.05).Compared with the NC group,the killing rate and the contents of TNF-αand IFN-γin the culture medium in the phloretin group,NK group,and NK+phloretin group increased in turn(F=330.181,288.215,336.648,all P<0.05).Conclusion Phloretin could enhance the killing activity of NK cells on GC cells,which might be related to destroying GC cell DNA,increasing the level of NKG2D ligands in the cell membrane,and promoting the

关 键 词：根皮素 自然杀伤细胞 胃癌 SGC-7901细胞 杀伤作用 

分 类 号：R735.2[医药卫生—肿瘤]