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目的 :观察全反式维甲酸 (RA)诱导分化前后 ,人髓母细胞瘤细胞系Med 3对抗Fas抗体的反应特点并探讨其免疫治疗意义。方法 :使用 10 μmol L的RA诱导Med 3细胞分化并在处理当时或第 2 4、48、72、96h ,分别加入最终浓度为 2 5 ,5 0 ,10 0 ,2 0 0和 40 0ng ml的抗Fas抗体。结合各组细胞Fas表达形式 ,分析其形态学 ,细胞动力学 ,克隆形成能力和死亡特点。结果 :正常培养的Med 3细胞表达 30kD的可溶型Fas(sFas)。RA能促进Med 3分化和膜型Fas(mFas)表达 ,但不诱导凋亡。单纯抗Fas抗体处理对靶细胞无明显效应。RA处理的同时或不同时期加入抗体能在不同程度上诱发细胞凋亡 ;其中 ,以RA处理后第 48h加入 2 0 0ng ml抗体的凋亡诱导效果最佳。结论 :RA通过上调mFas表达提高髓母细胞瘤细胞系的凋亡易感性。因此 ,RA与Fas相关性凋亡促进剂配伍可能有助于改善髓母细胞瘤的临床免疫和 或药物治疗现状。
OBJECTIVE: To observe the response of medullary cell line Med 3 to anti-Fas antibody before and after induced by all-trans retinoic acid (RA) and to explore its immunotherapy significance. METHODS: Med 3 cells were induced to differentiate with 10 μmol L of RA and were treated with either 20, 40, 50, 100, 200 and 40 ng of Anti-Fas antibody. Combined with the expression of Fas in each group of cells, the morphology, cell kinetics, clonogenic capacity and death characteristics were analyzed. Results: Normally cultured Med 3 cells express 30 kD of soluble Fas (sFas). RA promoted Med 3 differentiation and membrane Fas (mFas) expression but did not induce apoptosis. Anti-Fas antibody alone had no significant effect on target cells. RA at the same time or different times can induce apoptosis to varying degrees; Among them, the apoptosis induced by adding 200ng of antibody at 48h after RA treatment is the best. Conclusion: RA improves the susceptibility to apoptosis of medulloblastoma cell lines by up-regulating mFas expression. Therefore, the combination of RA and Fas-related apoptosis enhancers may help to improve the clinical immunology and / or drug therapy status of medulloblastoma.