摘要
目的观察Fas/FasL在绒毛膜癌细胞JEG-3、BeWo和Jurkat中的表达情况,探讨Fas/FasL与人绒癌细胞免疫逃逸的机制。方法免疫荧光染色法、RT-PCR、免疫印迹检测fas/fasl的表达。台盼蓝细胞拒染、四甲基偶氮唑蓝(MTT)法检测、流式细胞计数法检测绒癌细胞与Jurkat细胞共培养后对Jurkat细胞的抑制情况。结果①发现绒癌细胞上均有Fas及FasL的表达;②与Jurkat细胞相比,在基因和蛋白水平,绒癌细胞中Fas表达低下,而FasL表达升高。③绒癌细胞与Jurkat细胞共培养可诱导Jurkat细胞存活率下降(P<0.05),随共培养时间延长Jurkat细胞凋亡率升高(P<0.05);④用中和抗体NOK-2封闭细胞表面FasL后,Jurkat细胞的凋亡率从(17.86±5.59)%下降到(6.88±1.05)%。结论与Jurkat细胞相比绒癌细胞Fas表达下调而FasL的表达升高,证实Fas/FasL系统表达异常后可以使绒癌细胞逃离免疫监视。
【Objective】 To investigate the hitherto undescribed expression of Fas and FasL in the immortalized human choriocarcinoma cell lines BeWo and NJ-G,and to confirm the mechanism of T cell apoptosis induced by co-culture of choriocarcinoma cell with Jurkat.【Methods】 Receptor and ligand expression was demonstrated using specific antibodies and multiple techniques including confocal immunofluorescence microscopy,flow cytometry,immunoblots,MTT,Trypan blue dye exclusion assay and reverse transcription polymerase chain reaction（RT-PCR）.【Results】 ①Fas and FasL were expressed both in 2 human choriocarcinoma cell lines.②The expression of Fas in the level of both gene and protein,was lower in choriocarcinoma cells than that in Jurkat cells,while that of FasL was higher.③The choriocarcinoma cell induced the apoptosis of Jurkat cells in co-culture experiments.The apoptosis rate was increased with time.④The apoptosis rate of Jurkat cells in co-culture assays was decreased from（17.86±5.59）% to（6.88 ±1.05）% after FasL having been blocked with FasL neutralizing antibody NOK-2.【Conclusions】 Choriocarcinoma cells evade immune attack by down regulating the Fas receptor and killing lymphocytes through expression of FasL.Fas/ FasL pathway is one of the mechanisms of immune escape in choriocarcinoma cell cells and may be a new target point of immunological therapy.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2010年第21期3230-3235,共6页
China Journal of Modern Medicine
