曲古菌素A诱导Raji细胞和HL-60细胞表达共刺激分子CD80、CD86及其与免疫的关系

Expression of costimulatory molecular CD80 and CD86 in Raji cells and HL-60 cells induced by trichostatin A and the relationship with immunity

目的 研究曲古菌素A（TSA）诱导Raji细胞和HL-60细胞表达共刺激分子CD80和CD86及其与免疫应答的关系.方法 采用MTT法观察不同浓度TSA对Raji细胞和HL-60细胞的抑制作用,流式细胞仪检测150 nmol/L TSA作用Raji细胞和HL-60细胞的细胞活力及表达共刺激分子CD80和CD86,RT-PCR分析150 nmol/LTSA作用Raji细胞和HL-60细胞CD80和CD86 mRNA表达情况.结果 TSA对Raji细胞和HL-60细胞的抑制作用具有时间和剂量依赖性,流式细胞仪检测Raji细胞培养12、24、48 h表达CD80分别为（76.2±4.6）％、（78.7±6.9）％、（79.2±3.4）％,CD86表达极低,150 nmol/L TSA作用Raji细胞12、24、48 h后,可提高Raji细胞表达CD80,分别为[（82.4±7.2）％,t =2.56,P =0.047]、[（84.8土5.6）％,t=2.57,P=0.042]、[（93.9±8.7）％,t=2.67,P=0.038],CD86不上调;RT-PCR检测Raji细胞表达CD80 mRNA对照组条带亮度为0.45±0.32,150 nmol/L TSA作用Raji细胞12、24、48 h后表达CD80 mRNA增强,分别为（0.49±0.22,t=2.45,P =0.047）,（0.76 ±0.33,t=3.67,P=0.0071）,（0.85±0.34,t=4.25.P=0.0048）.CD86 mRNA不升高.流式细胞仪检测HL-60细胞12、24、48 h表达CD86分别为（28.8±5.3）％、（29.6±6.3）％、（29.2±1.4）％,CD80表达极低,150 nmol/LTSA作用HL-60细胞12、24、48 h后,提高HL-60细胞表达CD86,分别为[（38.5±4.3）％,t=2.87,P=0.037]、[（42.9±7.1）％,t=3.01,P=0.032]、[（61.4±9.7）％,t=4.56,P=0.0076],CD80不上调.RT-PCR检测HL-60细胞表达CD86 mRNA对照组为0.52 ±0.16,150 nmol/L TSA作用HL-60细胞12、24、48 h后表达CD86mRNA增强,分别为（0.63±0.45,t=2.67,P=0.042）,（0.75 ±0.61,t=3.97,P=0.0078）,（0.92±0.36,t=5.01,P=0.0032）,CD80 mRNA不升高.结论 TSA可诱导Raji细胞表达共刺激分子CD80及诱导HL-60细胞表达共刺激分子CD86,促进细胞的免疫应答,为抗恶性血液病提供了一个新的免疫治疗方法.

Objective To investigate trichostatin A （TSA）-induced costimulatory molecular C D80 and CD86 expressions of Raji and HL-60 cells as well as the relationship with immune response.Methods The proliferative activities of Raji cells and HL-60 cells were assessed by morphology and methyl thiazolyl tetrazolium （MTT） assay.The expressions of CD80 and CD86 were confirmed,and cell viability was examined by flow cytometer after 150 nmol/L TSA treatment.The expression status of CD80 mRNA and CD86 mRNA was analyzed by reverse transcription polymerase chain reaction （RT-PCR）.Results A time-and dose-dependent inhibition was detected in Raji cells and HL-60 cells treated with TSA.The expression of CD80 in Raji cells for 12,24,and 48 h was （76.2 ± 4.6） %,（78.7 ± 6.9） %,and （79.2 ± 3.4） %,respectively.After TSA （150 nmol/L） treatment for 12h,24h,and 48 h,the expression of CD80 was [（82.4±7.2）%,t =2.56,P =0.47],[（84.8 ±5.6）%,t =2.57,P =0.42],and [（93.9 ± 8.7） %,t =2.67,P =0.38] in Raji cells,respectively.The CD80 mRNA expression was 0.45 ± 0.32 in control group.After TSA （150 nmol/L） treatment in Raji cells for 12,24,and 48h,the CD80 mRNA expression was （0.49 ± 0.22,t =2.45,P =0.47）,（0.76 ± 0.33,t =3.67,P =0.0071）,and （0.85 ± 0.34,t =4.25.P =0.0048）,respectively.The expression of CD86 in HL-60 cells for 12h,24h,and 48 h was （28.8 ± 5.3） %,（29.6 ± 6.3） %,and （29.2 ± 1.4） %.After TSA （150 nmol/L） treatment for 12,24,and48 h,the expression of CD86 was [（38.5 ±4.3）%,t =2.87,P =0.37],[（42.9 ± 7.1） %,t =3.01,P =0.32],and [（61.4 ± 9.7） %,t =4.56,P =0.0076],respectively.The CD86 mRNA expression was 0.52 ±0.16 in control group.After TSA （150 nmol/L） treatment in HL-60 cells for 12h,24h,and 48 h,the CD86 mRNA expression was （0.63 ± 0.45,t =2.67,P =0.42）,（0.75 ± 0.61,t =3.97,P =0.0078）,and （0.92 ±0.36,t =5.01,P =0.0032）,respectively.Conclusions TSA induces costimulatory molecular CD80 and CD86 expression of Raji and HL-60 cells,improves immune response of cells,and provides a novel immunotherapy of ant-malignancy hematological disease.