摘要
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 mechanism of human enterovirus 71（ EV71） entering human neuroblastoma SK-N-SH cells. Methods After the SK-N-SH cells were pretreated with chlorpromazine（ CPZ） or nystatin（ NT）,real-time quantitative PCR（ qRT-PCR） was employed to measure EV71 mRNA level,and indirect immunofluorescence microscopy was used to detect the expression level of viral protein 1（ VP1） in the target cells. In order to reveal the colocalization of EV71 with clathrin,laser confocal microscopy was performed on the infected cells. Results CPZ could significantly inhibit EV71 mRNA level and the expression of VP1 in the target cells,while NT had no effect on EV71 infection. Confocal microscopy showed that EV71 was colocalize with clathrin. Conclusion EV71 infects human neuroblastoma SK-N-SH cel s by the clathrin-mediated endocytosis.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2017年第6期761-766,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
基金项目:重庆市科委(渝科发计字[2012]27号-cstc2012jjA10084)
