EZH2和H3K27me3的表达对食管癌细胞迁移和侵袭能力的影响
发布时间：2018-12-15 09:24
【摘要】：目的:通过转染Zeste同源物增强子2(enhancer of zeste homolog 2,EZH2)过表达或者敲低载体,探讨EZH2和Lys27位点三甲基化组蛋白H3(histone H3 methylated Lys27,H3K27me3)对食管麟状细胞癌(esophageal squamous cell cancer,ESCC)细胞迁移和侵袭能力的影响。方法:应用实时荧光定量PCR、Western blotting法检测ESCC细胞株KYSE30、KYSE170、TE1、Eca109中EZH2 mRNA水平,以及ESCC细胞过表达或者敲低EZH2对H3K27me3表达水平的影响。用划痕实验及Transwell侵袭实验分析过表达或者敲低EZH2后ESCC细胞的迁移侵袭能力。用实时荧光定量PCR法分析ESCC细胞过表达及敲低EZH2对MMPs mRNA水平的影响。结果:食管癌Eca109及TE1细胞中EZH2和H3K27me3 mRNA和蛋白水平明显高于KYSE30及KYSE170细胞(P0.05)。过表达EZH2的食管癌KYSE30及KYSE170细胞H3K27me3蛋白的表达水平显著升高(P0.05),敲低EZH2后Eca109及TE1细胞H3K27me3蛋白的表达水平明显降低(P0.05)。过表达EZH2后,KYSE30及KYSE170细胞的穿膜数目明显增多[(281.33±4.10)、(241.67±4.04)vs(132.00±4.00)、(105.33±3.51)个,均P0.05]、迁移距离明显增大[(63.6±1.2)、(62.5±2.5)vs(23.0±2.3)、(21.2±1.0)μm,P0.05]。敲低EZH2后Eca109及TE1细胞的穿膜数目显著减少(均P0.05),转染sh EZH2后Eca109及TE1细胞迁移的距离明显减小(均P0.05)。结论:EZH2可增加靶基因启动子上组蛋白H3第27位赖氨酸的三甲基化,并增强ESCC细胞的迁移和侵袭能力。
[Abstract]:Objective: to investigate the effect of EZH2 and Lys27 site trimethylated histone H3 (histone H3 methylated Lys27,H3K27me3) on (esophageal squamous cell cancer, in esophageal carcinoma by overexpression or knockdown of Zeste congener 2 (enhancer of zeste homolog 2 / EZH2). The effect of ESCC) on cell migration and invasion. Methods: the level of EZH2 mRNA in ESCC cell line KYSE30,KYSE170,TE1,Eca109 was detected by real-time fluorescence quantitative PCR,Western blotting, and the effect of ESCC cell over-expression or knockdown on H3K27me3 expression was evaluated. The migration and invasion ability of ESCC cells was analyzed by scratch test and Transwell invasion assay. The effect of over-expression of ESCC cells and knockout of EZH2 on MMPs mRNA level was analyzed by real-time fluorescence quantitative PCR. Results: the levels of EZH2, H3K27me3 mRNA and protein in Eca109 and TE1 cells were significantly higher than those in KYSE30 and KYSE170 cells (P0.05). The expression of H3K27me3 protein in KYSE30 and KYSE170 cells of esophageal carcinoma with overexpression of EZH2 was significantly increased (P0.05), and H3K27me3 protein expression in Eca109 and TE1 cells was significantly decreased after knocking down EZH2 (P0.05). After overexpression of EZH2, the number of perforating membrane of KYSE30 and KYSE170 cells increased significantly [(281.33 卤4.10), (卤4.04) vs (132.00 卤4.00), (105.33 卤3.51, P 0.05), respectively], and the migration distance was significantly increased [(63.6 卤1.2)]. (62.5 卤2.5) vs (23.0 卤2.3), (21.2 卤1.0 渭 m P 0.05). The transmembrane number of Eca109 and TE1 cells decreased significantly after EZH2 knockout (P0.05), and the migration distance of Eca109 and TE1 cells decreased significantly after sh EZH2 transfection (P0.05). Conclusion: EZH2 can increase the trimethylation of lysine on histone H3 on target gene promoter and enhance the migration and invasion of ESCC cells.
【作者单位】： 河北医科大学第四医院科研中心;
【基金】：河北省科技支撑计划资助项目(No.14277732D,No.152777184)~~
【分类号】：R735.1
[Abstract]:Objective: to investigate the effect of EZH2 and Lys27 site trimethylated histone H3 (histone H3 methylated Lys27,H3K27me3) on (esophageal squamous cell cancer, in esophageal carcinoma by overexpression or knockdown of Zeste congener 2 (enhancer of zeste homolog 2 / EZH2). The effect of ESCC) on cell migration and invasion. Methods: the level of EZH2 mRNA in ESCC cell line KYSE30,KYSE170,TE1,Eca109 was detected by real-time fluorescence quantitative PCR,Western blotting, and the effect of ESCC cell over-expression or knockdown on H3K27me3 expression was evaluated. The migration and invasion ability of ESCC cells was analyzed by scratch test and Transwell invasion assay. The effect of over-expression of ESCC cells and knockout of EZH2 on MMPs mRNA level was analyzed by real-time fluorescence quantitative PCR. Results: the levels of EZH2, H3K27me3 mRNA and protein in Eca109 and TE1 cells were significantly higher than those in KYSE30 and KYSE170 cells (P0.05). The expression of H3K27me3 protein in KYSE30 and KYSE170 cells of esophageal carcinoma with overexpression of EZH2 was significantly increased (P0.05), and H3K27me3 protein expression in Eca109 and TE1 cells was significantly decreased after knocking down EZH2 (P0.05). After overexpression of EZH2, the number of perforating membrane of KYSE30 and KYSE170 cells increased significantly [(281.33 卤4.10), (卤4.04) vs (132.00 卤4.00), (105.33 卤3.51, P 0.05), respectively], and the migration distance was significantly increased [(63.6 卤1.2)]. (62.5 卤2.5) vs (23.0 卤2.3), (21.2 卤1.0 渭 m P 0.05). The transmembrane number of Eca109 and TE1 cells decreased significantly after EZH2 knockout (P0.05), and the migration distance of Eca109 and TE1 cells decreased significantly after sh EZH2 transfection (P0.05). Conclusion: EZH2 can increase the trimethylation of lysine on histone H3 on target gene promoter and enhance the migration and invasion of ESCC cells.
【作者单位】： 河北医科大学第四医院科研中心;
【基金】：河北省科技支撑计划资助项目(No.14277732D,No.152777184)~~
【分类号】：R735.1
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