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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(P<0.05)。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(P<0.05),敲低EZH2后Eca109及TE1细胞H3K27me3蛋白的表达水平明显降低(P<0.05)。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P<0.05]、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,P<0.05]。敲低EZH2后Eca109及TE1细胞的穿膜数目显著减少(均P<0.05),转染sh EZH2后Eca109及TE1细胞迁移的距离明显减小(均P<0.05)。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。
OBJECTIVE: To investigate the effect of histone H3 methylated Lys27 (H3K27me3) on EZH2 and Lys27 sites by transfection with the enhancer of zeste homolog 2 (EZH2) overexpression or knockdown vector Effect of esophageal squamous cell carcinoma (ESCC) cell migration and invasion. Methods: The expression of EZH2 mRNA in ESCC cell lines KYSE30, KYSE170, TE1 and Eca109 was detected by real-time fluorescence quantitative PCR and Western blotting, and the effect of ESCC cell overexpression or knockdown of EZH2 on H3K27me3 expression was analyzed. Scratch assay and Transwell invasion assay were used to analyze the migration and invasion ability of ESCC cells after overexpression or knockdown of EZH2. The effect of EZH2 on the expression of MMPs mRNA in ESCC cells was analyzed by real-time fluorescence quantitative PCR. Results: The mRNA and protein levels of EZH2 and H3K27me3 in esophageal cancer Eca109 and TE1 cells were significantly higher than those in KYSE30 and KYSE170 cells (P <0.05). The expression of H3K27me3 was significantly increased in EZH2-overexpressing esophageal cancer KYSE30 and KYSE170 cells (P <0.05). The knockdown of EZH2 significantly decreased the expression of H3K27me3 in Eca109 and TE1 cells (P <0.05). The number of transmembrane of KYSE30 and KYSE170 cells was significantly increased after overexpression of EZH2 [(281.33 ± 4.10) vs (241.67 ± 4.04) vs (132.00 ± 4.00), (105.33 ± 3.51), both P <0.05] (63.6 ± 1.2), (62.5 ± 2.5) vs (23.0 ± 2.3), (21.2 ± 1.0) μm respectively, P <0.05]. The number of transmembrane of Eca109 and TE1 cells was significantly decreased after knockdown of EZH2 (both P <0.05), and the migration distance of Eca109 and TE1 cells was significantly decreased after shEZH2 transfection (all P <0.05). CONCLUSION: EZH2 can increase the methylation of lysine 27 of histone H3 on the target gene promoter and enhance the migration and invasion ability of ESCC cells.