PMA对Jurkat细胞株中可溶型分泌和膜型TRAIL表达的调节

目的:探讨乙酰肉豆蔻佛波酯(PMA)对Jurkat细胞可溶型TNF相关的凋亡诱导配体(solubletumornecrosisfac tor-relatedapoptosis inducingigand,sTRAIL)分泌和膜型TRAIL(membraneboundTRAIL,mTRAIL)表达的调节,以及二者的细胞毒作用。方法:分别采用ELISA及间接免疫荧光染色和流式细胞术分析,检测sTRAIL的分泌和mTRAIL的表达水平;用4h51Cr释放试验检测sTRAIL和mTRAIL对靶细胞Raji的细胞毒作用。结果:PMA刺激24h后jurkat细胞sTRAIL分泌和mTRAIL的表达均增加,sTRAIL分泌在刺激后48h达峰值,mTRAIL表达峰值在60h。两型TRAIL都具有对Raji细胞的细胞毒作用。结论:PKC活化剂PMA可上调Jurkat细胞sTRAIL分泌和mTRAIL的表达,两型TRAIL分子都具有杀伤靶细胞的细胞毒作用。

Regulation of soluble TRAIL and membrane TRAIL in Jurkat cells by PMA

AIM: To investigate the regulation of soluble and membrane bound TNF-ralated apoptosis-inducing ligand (TRAIL) in Jurkt cells by phorbol myristic acctate (PMA), and the cytotoxicity of the two forms of TRAIL. METHODS: Jurkat cells were cultured in the presence of 40 ng/mL PMA for different time. The production of sTRAIL was determined by ELISA, and expression of mTRAIL was analyzed by indirect fluorescence staining and flow cytometry analysis. The cytotoxicites of sTRAIL and mTRAIL were detected by ~(51)Cr release assay, in which DR4/DR5-expressing Raji cells were employed as target cells. RESULTS: The expression of both sTRAIL and mTRAIL in Jurkat cells were upregulated by PMA. The level of sTRAIL in supernatant from PMA-stimulated Jurkat cell culture increased and reached peak at 48 hours after PMA treatment, whereas expression peak of mTRAIL was at 60 hours. Both sTRAIL and mTRAIL exhibited cytotoxicity against Raji cells. CONCLUSION: PMA, a PKC activator, can upregulate the expression of both sTRAIL and mTRAIL in Jurkat cells, and the two forms of TRAIL have cytotoxic activity.