HDAC2 sumo-E3连接酶活性在DLD1细胞迁移和增殖中的作用  

Effect of HDAC2 Sumo-E3 Ligase Segment on the Proliferation and Migration of DLD1 Cells

作　　者：马琳琳 李玲 荆昭 李莹莹 郭韡[2] 胡雪停 邢伟[2] 徐祥 MA Lin-lin;LI Ling;JING Zhao;LI Ying-ying;GUO Wei;HU Xue-ting;XING Wei;XU Xiang(Qingdao University School of Basic Medical Sciences,Qingdao,Shandong,266071,China;State Key laboratory of Trauma,Burns and Combined Injury,Institute of Surgery Research,Daping Hospital,affiliated to the Third Military Medical University,Chongqing,400012,China)

机构地区：[1]青岛大学医学部基础医学院,山东青岛266071 [2]第三军医大学第三附属医院野战外科研究所创伤烧伤与复合伤国家重点实验室,重庆400012

出　　处：《现代生物医学进展》2018年第10期1854-1859,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金青年基金项目(81502434);重庆市基础与前沿研究计划一般项目(cstc2015jcyj A10078)

摘　　要：目的:探讨HDAC2 sumo-E3连接酶对DLD1细胞迁移与增殖的影响和可能机制。方法:全基因合成HDAC2 sumo-E3连接酶突变体HDAC2(325-488),并构建其重组慢病毒表达载体;感染DLD1hdac2-/-细胞(无内源性HDAC2表达),筛选稳定表HDAC2(325-488)的细胞克隆DLD1h325-488;RTCA实时细胞分析仪、Trans-well等检测DLD1h325-488稳定细胞的增殖和迁移能力;免疫印迹、q RT-PCR检测DLD1h325-488稳定细胞增殖和迁移相关基因的表达。结果:构建了稳定表达HDAC2 sumo-E3连接酶突变体HDAC2(325-488)的细胞株DLD1h325-488,并获得稳定表达HDAC2 sumo-E3连接酶突变体HDAC2(325-488)的DLD1h325-488细胞。与对照组细胞相比,DLD1h325-488细胞增殖活性无显著变化,但是细胞的迁移能力和迁移相关蛋白MMP14的表达显著上升。结论:HDAC2 sumo-E3连接酶可能通过上调mmp14的表达而促进DLD1细胞的迁移。Objective： To investigate the effect of HDAC2 sumo-E3 ligase segment on proliferation and migration of DLD1 cells and the potential mechanisms.Methods： The coding sequence of the sumo-E3 ligase segment of HDAC2（325-488 aa） was chemically synthesized and ligated into a recombinant lentiviral vector.The lentiviral vector was packaged into lentiviral particles and used to infect HDAC2-knockout DLD1 cells（DLD1-HDAC2-/-）,and stably transfected cells were selected and named DLD1 h325-488.Then,proliferation and migration of the DLD1 h325-488 cells were determined by RTCA and transwell assay; and changes in expression of proliferation-and migration-related genes were determined by q RT-PCR and immune-blotting.Results： Stable DLD1 h325-488 cell line was successfully obtained.Compared to the control cells,and DLD1 h325-488 cells stably expressing HDAC2 sumo-E3 ligase mutant.HDAC2（325-488）.DLD1 h325-488 cells showed no significant changes in proliferation,but significantly increased migration,which was accompanied by increase expression of the migration-related MMP14.Conclusion： HDAC2 sumo-E3 ligase segment may promote the migration of DLD1 cells by upregulating MMP14 expression.

关 键 词：HDAC2 SUMO 迁移 MMP14 

分 类 号：Q55[生物学—生物化学] Q78