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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 and FasL in human laryngeal carcinoma Hep-2 cells and to explore the role of Fas / FasL pathway in immune escape of human laryngeal carcinoma. Methods: The mRNA and protein expression of Fas / FasL on Hep-2 cells were detected by RT-PCR and flow cytometry. Hep-2 cells were co-cultured with Jurkat cells. MTT assay was used to characterize the growth curve of Jurkat cells. Flow cytometry Hurchst staining and fluorescence microscopy were used to observe the apoptosis of Jurkat cells. Results: The average fluorescence intensity of Fas and FasL on Hep-2 cells by flow cytometry was 32.91 ± 5.6 and 25.57 ± 7.1, respectively. Hep-2 cells (density 1 × 109 / L) were co-cultured with Jurkat cells (density 1 × 108 / L, 5 × 108 / L) for 24 h. The apoptosis rate of Jurkat cells by flow cytometry was (38.95 ± 0.11)% and (13.28 ± 0.14)%, respectively. The apoptosis rate of Jurkat cells cultured alone was (7.53 ± 0.17)%. MTT colorimetric assay showed that Jurkat cell growth was significantly inhibited after co-culture. When the neutralizing antibody was added into the coculture system, the apoptosis rate of Jurkat cells was significantly decreased (P <0.01). Conclusion: FasL expressed on the surface of human laryngeal carcinoma cell line Hep-2 can evade host immune surveillance by inducing apoptosis of activated T lymphocyte Jurkat cells.