白细胞介素21及白细胞介素12单用或联合应用对子宫内膜癌患者外周血单核细胞抗肿瘤活性的影响  

Effect of Interleukin 21 and/or Interleukin 12 on the Antitumor activity of Peripheral Blood Mononuclear Cells in Patients with Endometrial Cancer

作　　者：田永菊[1] 崔保霞[1] 马道新[2] 张妍[1,3] 侯菲[1] 张文静[1] 

机构地区：[1]山东大学齐鲁医院妇产科,济南250012 [2]山东大学齐鲁医院血液病研究室,济南250012 [3]山东省潍坊市人民医院妇产科,山东潍坊261041

出　　处：《中国医学科学院学报》2011年第3期292-298,共7页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(30400475);山东省优秀中青年科学家科研奖励基金计划(2006BS03060);留学回国科研基金~~

摘　　要：目的观察白细胞介素(IL)21及IL12单用或联合应用对子宫内膜癌患者外周血单核细胞(PBMCs)抗肿瘤活性的影响。方法体外分离子宫内膜癌患者外周血中的PBMCs,用低浓度IL2维持培养,经不同的刺激条件(对照组、抗IL21组、IL21组、IL12组和IL21+IL12联合组)诱导培养72 h后,采用乳酸脱氢酶(LDH)释放法检测PBMCs的细胞毒活性,流式细胞术检测CD4+CD25+FOXP3+T调节性细胞(Treg细胞)和CD4+IL17A+T辅助17细胞(Th17细胞)的含量,细胞活性检测试剂-8(CCK-8)法检测PBMCs的增殖能力,流式细胞术检测PBMCs的凋亡情况。结果与对照组相比,IL21及IL12可显著增强PBMCs细胞毒活性,IL21+IL12联合组的效果优于单用IL21组和IL12组。IL21及IL12可显著降低Treg细胞含量,抑制PBMCs凋亡。不同细胞因子处理组对Th17细胞的含量和PBMCs的增殖能力均无显著影响。结论 IL21及IL12可增强子宫内膜癌患者PBMCs的细胞毒活性,联合应用效果更明显。抑制Treg细胞的分化和PBMCs的凋亡可能为其机制之一,而与Th17细胞的分化无关。Objective To observe the role of interleukin（IL） 21 alone,IL12 alone,and IL21 plus IL12 for inducing the antitumor activity of peripheral blood mononuclear cells（PBMCs） in patients with endometrial cancer.Methods PBMCs were isolated from peripheral blood in patients with endometrial cancer in vitro,and kept the culture with low-level IL2.IL2-stimulated PBMCs were cocultured under different conditions（with anti-IL21 antibody,IL21 alone,IL12 alone,or IL21 plus IL12） for 72 h.The cytotoxicity of PBMCs was then examined by lactate dehydrogenase（LDH） released assay.CD4＋ CD25＋ FOXP3＋T regulatory（Treg） cell and CD4＋ IL17A＋ T-helper（Th17） cell proportion were determined with flow cytometry.Cell proliferation and apoptosis were measured by cell counting kit-8（CCK-8）assay and flow cytometry,respectively.Results In comparison to control group,both IL21 and IL12 significantly enhanced the cytotoxicity of PBMCs.The IL21 plus IL12 group had superior effect to IL21 alone and IL12 alone.IL21 and IL12 significantly decreased the percentages of Treg cells and the rate of PBMCs apoptosis.IL21 or IL12 had no significant effect on the differentiation of Th17 cells and the proliferation of PBMCs.Conclusions IL21 and IL12 can enhance the cytotoxicity of PBMCs in patients with endometrial cancer,which can be further strengthened with treatment of IL21 plus IL12.Such effects may be achieved by inhibiting the differentiation of Treg cells and the apoptosis of PBMCs,but not by the differentiation of Th17 cell.

关 键 词：细胞因子 外周血单核细胞 细胞毒活性 TREG细胞 TH17细胞 

分 类 号：R737.33[医药卫生—肿瘤]