| 马琳琳,李 玲,荆 昭,李莹莹,郭 韡,胡雪停,邢 伟,徐 祥.HDAC2 sumo-E3连接酶活性在DLD1细胞迁移和增殖中的作用[J].,2018,(10):1854-1859 |
| 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 |
| 投稿时间：2017-10-18 修订日期：2017-11-10 |
| DOI：10.13241/j.cnki.pmb.2018.10.010 |
| 中文关键词: HDAC2 SUMO 迁移 MMP14 |
| 英文关键词: HDAC2 SUMO Migration MMP14 |
| 基金项目: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(cstc2015jcyjA10078) |
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| 中文摘要: |
| 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、qRT-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。 |
| 英文摘要: |
| ABSTRACT 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 (325-488aa) 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 DLD1h325-488. Then, prolifer- ation and migration of the DLD1h325-488 cells were determined by RTCA and transwell assay; and changes in expression of prolifera- tion- and migration-related genes were determined by qRT-PCR and immune-blotting. Results: Stable DLD1h325-488 cell line was suc- cessfully obtained. Compared to the control cells, and DLD1h325-488 cells stably expressing HDAC2 sumo-E3 ligase mutant. HDAC2 (325-488). DLD1h325-488 cells showed no significant changes in proliferation, but significantly increased migration, which was accom- panied by increase expression of the migration-related MMP14. Conclusion: HDAC2 sumo-E3 ligase segment may promote the migra- tion of DLD1 cells by upregulating MMP14 expression. |
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