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

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.