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目的:研究IL-16对人白血病细胞系MT2细胞增殖抑制和凋亡的影响,为进一步研究IL-16功能奠定基础。方法:IL-16腺病毒载体转染MT2细胞后,不同时间收获细胞,通过MTT、流式细胞术(FCM)、测定线粒体膜电位和单细胞凝胶电泳等方法,证实MT2细胞增殖抑制和凋亡。结果:用MTT法绘制生长曲线,观察到IL-16腺病毒载体转染的MT2细胞增殖受到明显抑制;DNA片段分析在转染6h后出现凋亡片段;进一步用FCM检测细胞周期停滞于G1期,并且出现细胞凋亡;单细胞凝胶电泳发现细胞12h后有明显凋亡DNA拖尾形成;IL-16对细胞线粒体膜电位的影响24h内逐渐增强,预示细胞凋亡;免疫荧光发现12h自噬相关蛋白LC3,即可在细胞膜上表达,自噬已经开始出现。结论:重组人IL-16腺病毒载体转染MT2细胞,24h内即可诱导MT2细胞发生细胞增殖抑制并且出现凋亡,为进一步研究IL-16的功能奠定了基础。
Objective: To study the effect of IL-16 on the proliferation and apoptosis of human leukemia cell line MT2 cells, and to lay the foundation for further study on the function of IL-16. Methods: After transfection of MT2 cells with IL-16 adenovirus vector, cells were harvested at different times. MTT, flow cytometry (FCM), mitochondrial membrane potential and single cell gel electrophoresis were used to confirm the inhibition of MT2 cell proliferation and apoptosis Death. Results: The growth curve was drawn by MTT method. The proliferation of MT2 cells transfected with IL-16 adenovirus vector was significantly inhibited. The apoptotic fragments were observed 6 h after transfection with DNA fragment. The cell cycle arrest was further detected by FCM in G1 phase , And the apoptosis appeared; single cell gel electrophoresis found that the cells after 12h apparent apoptosis of DNA tail formation; IL-16 on mitochondrial membrane potential gradually increased within 24h, indicating apoptosis; immunofluorescence 12h LC3, which is expressed on the cell membrane, autophagy has begun to appear. CONCLUSION: MT2 cells transfected with recombinant human IL-16 adenovirus vector can induce cell proliferation inhibition and apoptosis in MT2 cells within 24h, which lays the foundation for further study on the function of IL-16.