FITC标记重组灵芝免疫调节蛋白(rLz-8)在NB4细胞中的动态定位

Dynamic Observation of Cellular Localization of Fluorescein Isothiocyanate Labeled Recombinant Ganoderma Lucidum Immunoregulatory Protein(rLz-8) in NB4 APL Cell

实验发现,重组灵芝免疫调节蛋白(rLz-8)可直接杀伤人急性早幼粒细胞白血病细胞株NB4.利用异硫氰酸荧光素(FITC)标记rLz-8,其相关的活性实验结果和晶体结构分析都表明,FITC没有影响rLz-8已知生物学功能.通过激光共聚焦显微镜观察FITC-rLz-8在NB4细胞内的动态过程发现,FITC-rLz-8可识别细胞膜上的受体,并可进入细胞质,并最终富集在细胞核区域内.Annexin V-FITC双染检测结果显示,rLz-8对NB4细胞杀伤作用的可能机制是对NB4细胞凋亡的诱导作用,在一定浓度范围内,剂量与凋亡诱导率成正相关.因此rLz-8能够诱导肿瘤细胞NB4发生凋亡的亚细胞学机制可定位在细胞核上.

NB4 APL cell was induced apoptosis by Recombinant Ganoderma lucidum immunoregulatory pro- tein（rLz-8） for the first time in this study. The dynamic cellular localization of FITC-rLz-8 was observed by laser scanning confocal microscope LSCM： identifying the receptors of cell membrane, entering into cell, gathering on nucleus finally. Annexin V-FITC kit was used for measuring apoptosis of NB4 by rLz-8. Results show that the effects of the dose group（0.5 μg/mL） is close to the one of positive control（As203）. All data show that apoptosis inducing activity of rLz-8 in HL60 cells and changes of cell morphology and Ca^2＋ level in cytosol are probably interrelate with rLz-8＇s subcellular localization.