Plk1在猪胎儿成纤维细胞有丝分裂进程中的亚细胞定位和功能研究  

Subcellular localization and functional study of Plk1 in the mitosis process of pig fetal fibroblasts

作　　者：林德凤 董妍 石凤垚 曹伟 苗懋东 芮荣[1] 剧世强[1] LIN Defeng;DONG Yan;SHI Fengyao;CAO Wei;MIAO Maodong;RUI Rong;JU Shiqiang(College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China)

机构地区：[1]南京农业大学动物医学院

出　　处：《畜牧与兽医》2018年第7期51-56,共6页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31572589,31201967);江苏高校优势学科建设工程资助项目

摘　　要：本研究旨在探究Plk1在猪胎儿成纤维细胞中的亚细胞定位及其作用。通过间接免疫荧光技术检测了Plk1在猪胎儿成纤维细胞有丝分裂进程中的亚细胞定位,并通过Plk1特异性抑制剂GSK461364处理进一步分析Plk1在细胞有丝分裂进程中的潜在作用。结果显示：Plk1在猪胎儿成纤维细胞有丝分裂各个时期都有分布,且在细胞核周围有相对较高水平的Plk1表达;经GSK461364处理后,细胞活力显著降低,细胞周期进程阻滞在G2/M期,出现细胞核碎裂,并伴随着α-微管蛋白结构紊乱。研究表明：Plk1参与了调控猪胎儿成纤维细胞有丝分裂的进程,并与G2/M期的细胞核分裂和微管组装密切相关。This study was conducted to address the localization and possible role of Plk1 in porcine fetal fibroblasts during mitosis. Indirect immunofluorescence was used to determine the dynamic subcellular localization of Plk1 in porcine fetal fibroblasts,and a specific Plk1 inhibitor,GSK461364,was employed to evaluate the potential role of Plk1 during the mitotic progression. The results showed that microfilaments and microtubules exhibited a dynamic distribution pattern,which were related to the cell cycle progression during mitotic division in porcine fetal fibroblasts. The signal of Plk1 could be continuously detected in porcine fetal fibroblasts undergoing mitosis,and a relatively higher level of Plk1 was observed around the nucleus during the mitotic division progression. GSK461364 treatment significantly reduced cell viability and proliferative capacity in a dose-dependent manner,with a higher proportion of Plk1-inhibited cells arrested in the G2/M stage. Further subcellular structure examination showed that Plk1 inhibition led to significantly higher percentages of the GSK461364-treated cells that exhibited fragmented nuclei,which was accompanied by the misarranged α-tubulin. It could be concluded that Plk1 was essential for the mitotic entry into porcine fetal fibroblasts,and the regulation was associated with the nuclear activities and microtubule organization during the G2/M stage.

关 键 词：猪胎儿成纤维细胞 PLK1 有丝分裂 细胞核 Α-微管蛋白 

分 类 号：S857.2[农业科学—临床兽医学]