FasL反义RNA抑制人T淋巴细胞致凋亡作用的研究

Reducing apoptosis-inducing capacity of the human T lymphocytes blocked by anti-sense RNA of FasL

目的 :探讨反义RNA抑制活化T淋巴细胞FasL的表达及其对Fas阳性细胞的致凋亡作用 ,为FasL反义RNA的应用提供实验依据。方法 :获得FasL反义RNA重组逆转录病毒 ,调整感染滴度后转染活化人外周血单个核细胞 (HPBMC)。采用流式细胞仪检测转染细胞表面FasL的表达以及对Fas+HL 60细胞的致凋亡能力。结果 :空病毒载体的重复转染上调活化HPBMCFasL的表达 ,但转染重组病毒能有效抑制活化HPBMC表达FasL ,表达率由13 93 %下降至 7 19% ;FACS检测HL 60细胞的凋亡显示 ,转染重组病毒使活化HPBMC对HL 60细胞的致凋亡作用明显下降 ,凋亡率由 3 3 48%降为 17 2 8%。结论 :FasL反义RNA能有效抑制活化淋巴细胞表达FasL及由其介导的凋亡作用 。

Objective To investigate whether the apoptosis inducing capacity of the activated T lymphocytes can be reduced by FasL anti sense RNA.Method Flowcytometry was carried to analyze the FasL expression of human peripheral blood mononuclear cells (HPBMC) transfected with recombinant retrovirus vector which was able to transcript anti sense RNA of FasL.After co incubated with the transfected HPBMC,the cellular apoptotic rate of Fas positive HL 60 cells was tested using FACS.Results The FasL expression level of the HPBMC transfected with recombinant retrovirus vector was down regulated significantly compared with mock vector(7.19% vs.13.93%).The cellular apoptotic rate of HL?60 cells co incubated with the HPBMC transfected with recombinant retrovirus vector was 17.28%,much lower than 33.48% of co incubated with the HPBMC transfected with mock vector.Conclusion FasL expression and apoptosis inducing capacity of human T lymphocytes can be effectively blocked by the anti sense RNA.