IL对NK细胞杀伤人骨髓瘤RPMI 8226细胞活性的影响及其机制  

Influence of interleukin in inducing the cytotoxicity of NK cells against human multiple myeloma RPMI 8226 cells and its mechanism

作　　者：韩露[1] 高全立[1] 宋永平[1] 周健[1] 

机构地区：[1]郑州大学附属肿瘤医院河南省肿瘤医院,郑州450008

出　　处：《山东医药》2016年第12期1-4,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81000921)

摘　　要：目的探讨IL对NK细胞杀伤人骨髓瘤RPMI 8226细胞活性的影响,并探讨其机制。方法取正常人外周血单个核细胞,随机分为6组,对照组以单纯培养基培养,IL-2组加入1 000 U/m L IL-2,IL-7组加入40 ng/m L IL-7,IL-21组加入20 ng/m L IL-21,IL-2+IL-7组分别加入1 000 U/m L IL-2和40 ng/m L IL-7,IL-2+IL-7+IL-21组分别加入1 000 U/m L IL-2、40 ng/m L IL-7和20 ng/m L IL-21,均培养5天。采用流式细胞术检测各组NK细胞比例及其表面激活性受体NKG2D、NKp46、NNKp30阳性表达细胞比例,RT-PCR法检测各组NK细胞NKG2D mRNA、NKP46 mRNA和NKP30 mRNA,CFSE/PI双染法检测各组NK细胞对RPMI 8226细胞的细胞毒效应(杀伤率)。结果除IL-7组外,其余各组NK细胞比例和细胞表面NKG2D阳性表达细胞比例均较对照组升高(P均<0.05)。各组NK细胞NKG2D mRNA、NKP46 mRNA和NKP30 mRNA表达量比较差异均无统计学意义(P均>0.05)。对照组NK细胞对RPMI 8226细胞的杀伤率为2.13%±0.42%、IL-2组为58.73%±1.80%、IL-7组为1.90%±0.60%、IL-21组为14.43%±1.22%、IL-2+IL-7组为34.27%±2.35%、IL-2+IL-7+IL-21组为37.47%±0.60%,各组与对照组比较P均<0.05。结论单因子IL-2、IL-21,双因子IL-2+IL-7和多因子IL-2+IL-7+IL-21均能提高NK细胞对人骨髓瘤RPMI 8226细胞的杀伤活性,以单因子IL-2作用效果最好;其作用机制可能为IL增加NK细胞数量和其表面活化性受体NKG2D的表达量。Objective To investigate the influence of interleukin（ IL） in inducing the cytotoxicity of natural killer（ NK） cells against the activity of human multiple myeloma RPMI 8226 and the correlated molecular mechanisms. Methods The human peripheral blood mononuclear cells（ PBMC） were randomly divided into 6 groups： the control group（ simple culture medium）,IL-2 group（ added with 1 000 U / m L IL-2）,IL-7 group（ 40 ng / m L IL-7）,IL-21 group（ 20 ng / m L IL-21）,IL-2 ＋ IL-7 group（ 1 000 U / m L IL-2 ＋ 40 ng / m L IL-7） and IL-2 ＋ IL-7 ＋ IL-21 group（ 1 000 U / m L IL-2,40 ng /m L IL-7 ＋ 20 ng / m L IL-21）. They were all cultured for 5 days. The proportion of NK cells and the expression of activated receptors（ NKG2 D,NKp46,NKp44 and NNKp30） were measured by flow cytometry. The expression of NKG2 D mRNA,NKP46 mRNA and NKP30 mRNA was measured by RT-PCR. Cytotoxicity of NK cells against RPMI8226 cells was analyzed by CFSE / PI double staining. Results Except the IL-7 group,the proportion of NK cells and the expression of NKG2 D in the other groups were increased as compared with those of the control group（ all P〈0. 05）. No significant difference in the gene expression of NKG2 D,NNKp46 and NKp30 was found among these groups（ all P〈0. 05）. The cellkilling rate of NK cells on RPMI8226 cells in the control group was 2. 13% ± 0. 42%,and that in the IL-2 group was58. 73% ± 1. 80%,the IL-7 group was 1. 90% ± 0. 60%,the IL-21 group was 14. 43% ± 1. 22%,the IL-2 ＋ IL-7 group was 34. 27% ± 2. 35%,and the IL-2 ＋ IL-7 ＋ IL-21 group was 37. 47% ± 0. 60%. Significant difference was found in the cell-killing rate between the control group and the other groups（ all P〈0. 05）. Conclusions The single factor IL-2,IL-21,double-factor IL-2 ＋ IL-7 and multi-factor IL-2 ＋ IL-7 ＋ IL-21 can enhance the cytotoxic effect of NK cells against RPMI 8226 cells,and the single factor IL-2 is the best. The increased number of NK cells and the expression of cell surface acti

关 键 词：多发性骨髓瘤 自然杀伤细胞 白细胞介素 天然细胞毒受体 NKG2D 

分 类 号：R733.3[医药卫生—肿瘤]