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本文探讨细胞表面β-1,4-半乳糖基转移酶-Ⅰ(β-1,4-galactosyltransferase-I,β-1,4-GalT-Ⅰ)在Jurkat细胞活化过程中的作用。采用植物凝集素(phytohemagglutinin,PHA)刺激Jurkat细胞,CCK-8法检测Jurkat细胞增殖;分别采用实时荧光定量PCR(RFQ-PCR)技术、Western blot技术及流式细胞术检测活化过程中的不同时间点β-1,4-GalT-Ⅰ在Jurkat细胞中的表达变化;采用免疫荧光细胞化学法和激光共聚焦技术观察Jurkat细胞表面β-1,4-GalT-Ⅰ的分布。Jurkat细胞经PHA刺激后增殖,在36 h达高峰;细胞表面β-1,4-GalT-Ⅰ的mRNA表达量、蛋白表达量以及平均荧光表达强度均较未活化状态增高;Jurkat细胞表面β-1,4-GalT-Ⅰ在未活化状态下的平均分布于细胞表面,活化后其发生重排和聚集。结果提示在T细胞活化过程中细胞表面β-1,4-GalT-Ⅰ分子的表达增高且发生了分布变化。
This article investigates the role of β-1,4-galactosyltransferase-I (β-1,4-GalT-I) in the activation of Jurkat cells. Jurkat cells were stimulated with phytohemagglutinin (PHA), and proliferation of Jurkat cells was detected by CCK-8 assay. Real-time quantitative PCR (RFQ-PCR), Western blot and flow cytometry Point β-1,4-GalT-Ⅰ in Jurkat cells. The distribution of β-1,4-GalT-Ⅰ on Jurkat cells was observed by immunofluorescence cytochemistry and confocal laser scanning microscope. Jurkat cells proliferated after being stimulated by PHA and peaked at 36 h. The mRNA expression, the protein expression level and the average fluorescence intensity of β-1,4-GalT-Ⅰ on the cell surface were all higher than those of the untreated Jurkat cells. 1,4-GalT-I in the non-activated state of the average distribution on the cell surface, activated rearrangement and aggregation occurs. The results suggest that the expression of β-1, 4-GalT-I molecules on the cell surface is increased and the distribution changes during T cell activation.