摘要
目的:观察人喉癌Hep-2细胞Fas和FasL的表达,探讨Fas/FasL途径在人喉癌细胞免疫逃避中的作用。方法:应用RT-PCR、流式细胞仪检测Hep-2细胞表面Fas/FasL的mRNA及蛋白表达;Hep-2细胞与Jurkat细胞共培养,MTT比色法描绘Jurkat细胞生长曲线,应用流式细胞术及Hoechst染色荧光显微镜观察及检测Jurkat细胞凋亡。结果:流式细胞仪检测Hep-2细胞表面Fas及FasL表达的平均荧光强度分别为32.91±5.6和25.57±7.1。Hep-2细胞(密度为1×109/L)分别与Jurkat细胞(密度为1×108/L、5×108/L)共培养24 h,流式细胞仪检测Jurkat细胞凋亡率为(38.95±0.11)%和(13.28±0.14)%,而Jurkat细胞单独培养的凋亡率为(7.53±0.17)%。MTT比色法检测显示共培养后Jurkat细胞生长受到明显抑制;当共培养体系中加入FasL中和性抗体后,Jurkat细胞凋亡率显著下降(P<0.01)。结论:人喉癌细胞株Hep-2表面表达的FasL通过诱导活化T淋巴细胞Jurkat细胞产生凋亡而使肿瘤细胞逃避宿主的免疫监视。
Objective： To observe the expression of Fas receptor and ligand in human laryngocarcinoma cell line, Hep-2 and to investigate the possible mechanism of immune escape through Fas/FasL pathway in Hep-2 cell. Method：The mRNA and protein expressions of Fas and FasL in Hep-2 cell were analysized by RT-PCR and flow cytometry （FCM）. Growth curve of Jurkat cell was drawed based on the results of MMT, and apoptosis of Jurkat cell were determined by FCM and Hoecbst 33342 staning after coeuhuring with Hep-2 cell. Result：The expressions of Fas and FasL in Hep-2 cell line were evaluated by flow cytometry and the mean fluorescence indensity were （32.91±5.6） and （25. 57±7. 1） respectively. After coincubaiton with Hep 2 cell （1 ×10^9/L）, the apoptosis rates of Jurkat cells were （38.95±0.11）% and （13.28±0.14）%, with planting concentration at 1 × 10^8/L and 5 ×10^8/L respectively. In contrast, the apoptosis rate of Jurkat cultured separatedly was （7.53 ± 0.17）%. The proliferation of Jurkat cell was obviously inhibited after cocuhure. However, the apoptosis rate was significantly decreased after adding neutralizing antibody of FasL. Conclusion： Laryngocarcinoma cell could induce apoptosis of T lympbocytes through Fas-FasL system, thus it provided a potential mecbansim to escape from immune surveillance of host.
出处
《临床耳鼻咽喉头颈外科杂志》
CAS
CSCD
北大核心
2008年第12期560-563,共4页
Journal of Clinical Otorhinolaryngology Head And Neck Surgery
关键词
喉肿瘤
T淋巴细胞
凋亡
免疫逃避
Laryngeal neoplasms
T lymphocytes
Apoptosis
Immune escape
