中国病理生理杂志2011，Vol.27Issue(2)：382-385,4.DOI:10.3969/j.issn.1000-4718.2011.02-033
机械牵张诱导THP-1细胞高迁移率族蛋白B1移位出核
Mechanical stretch induces translocation of high-mobility group box 1 protein in THP-1 cells
摘要
Abstract
AIM : To investigate the translocation of high - mobility group box 1 protein( HMCBI ) in THP - 1cells induced by mechanical stretch.METHODS: The vector that expressed the fusion protein of HMCB1 and enhanced green fluorescent protein( EGFP ) was constructed.The cDNA of HMGB1 and EGFP was subcloned into hemagglutinin ( HA ) - tagged vector pcDNA3 - HA by two - step methos.THP - 1 cells were transfected with pcDNA3 - HMGB1 - EGFP and exposed to cyclic mechanical stretch at 20％ elongation using Flexercell 4000T cell stretching unit.The translocation of HMGB1 in THP - 1 cells was ohserved under fluorescence microscope.RESULTS : The recombinant plasmid was verified by enzyme digestion.The green fluorescence accumulated in the nuclei of the cells, indicating that the fusion protein was highly expressed in THP - 1 cells and localized in the nuclei.Eighteen hours after mechanical stretch, the green fluorescence was observed in the cytoplasm.At the same time, the green fluorescence was still localized in the nuclei of the control cells treated without mechanical stretch.CONCLUSION : The HMGB1 - EGFP fusion protein is successfully and effectively expressed in THP - 1 cells.Mechanical stretch induces the translocation of HMGB1 protein from nucleus to cytoplasm.关键词
机械牵张/高迁移率族蛋白质类/THP-1细胞分类
医药卫生引用本文复制引用
丁宁,肖慧,许立新,佘守章..机械牵张诱导THP-1细胞高迁移率族蛋白B1移位出核[J].中国病理生理杂志,2011,27(2):382-385,4.基金项目
广东省科技计划资助项目(No.2010B031600011) （No.2010B031600011）
广东省医学科研基金资助项目(No.A2009497) （No.A2009497）
广州市医药卫生科技一般引导项目(No.2009-YB-021) （No.2009-YB-021）
广州市医药卫生科技重点资助项目(No.2008-ZDi-14) （No.2008-ZDi-14）
