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目的研究肠道病毒71型(EV71)入侵人神经母细胞瘤SK-N-SH细胞的内吞机制。方法利用氯丙嗪(CPZ)、制霉菌素(NT)分别预处理SK-N-SH细胞,实时定量PCR(qRT-PCR)检测靶细胞中EV71 mRNA水平,免疫荧光细胞化学染色检测靶细胞病毒蛋白1(VP1)蛋白表达水平;EV71感染SK-N-SH细胞后,激光共聚焦显微镜检测EV71与入侵途径标志分子网格蛋白的共定位。结果 CPZ能抑制靶细胞内EV71 mRNA和VP1蛋白的表达,而NT对其无影响;激光共聚焦显微镜发现EV71与网格蛋白共定位。结论 EV71是通过网格蛋白介导的内吞途径入侵人神经母细胞瘤SK-N-SH细胞。
Objective To study the endocytosis mechanism of enterovirus 71 (EV71) invader human neuroblastoma SK-N-SH cells. Methods SK-N-SH cells were pretreated with chlorpromazine (CPZ) and nystatin (NT), respectively. The levels of EV71 mRNA in target cells were detected by qRT-PCR and immunofluorescence cytochemistry Protein 1 (VP1) protein expression. EV71 infected SK-N-SH cells were examined by confocal laser scanning microscopy co-localization of EV71 and invasion molecular marker protein. Results CPZ could inhibit the expression of EV71 mRNA and VP1 protein in target cells, while NT had no effect on it. Laser confocal microscopy revealed that EV71 co-localized with clathrin. Conclusion EV71 invades human neuroblastoma SK-N-SH cells via the clathrin-mediated endocytosis pathway.