【摘要】：目的:通過(guò)轉(zhuǎn)染Zeste同源物增強(qiáng)子2(enhancer of zeste homolog 2,EZH2)過(guò)表達(dá)或者敲低載體,探討EZH2和Lys27位點(diǎn)三甲基化組蛋白H3(histone H3 methylated Lys27,H3K27me3)對(duì)食管麟狀細(xì)胞癌(esophageal squamous cell cancer,ESCC)細(xì)胞遷移和侵襲能力的影響。方法:應(yīng)用實(shí)時(shí)熒光定量PCR、Western blotting法檢測(cè)ESCC細(xì)胞株KYSE30、KYSE170、TE1、Eca109中EZH2 mRNA水平,以及ESCC細(xì)胞過(guò)表達(dá)或者敲低EZH2對(duì)H3K27me3表達(dá)水平的影響。用劃痕實(shí)驗(yàn)及Transwell侵襲實(shí)驗(yàn)分析過(guò)表達(dá)或者敲低EZH2后ESCC細(xì)胞的遷移侵襲能力。用實(shí)時(shí)熒光定量PCR法分析ESCC細(xì)胞過(guò)表達(dá)及敲低EZH2對(duì)MMPs mRNA水平的影響。結(jié)果:食管癌Eca109及TE1細(xì)胞中EZH2和H3K27me3 mRNA和蛋白水平明顯高于KYSE30及KYSE170細(xì)胞(P0.05)。過(guò)表達(dá)EZH2的食管癌KYSE30及KYSE170細(xì)胞H3K27me3蛋白的表達(dá)水平顯著升高(P0.05),敲低EZH2后Eca109及TE1細(xì)胞H3K27me3蛋白的表達(dá)水平明顯降低(P0.05)。過(guò)表達(dá)EZH2后,KYSE30及KYSE170細(xì)胞的穿膜數(shù)目明顯增多[(281.33±4.10)、(241.67±4.04)vs(132.00±4.00)、(105.33±3.51)個(gè),均P0.05]、遷移距離明顯增大[(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細(xì)胞的穿膜數(shù)目顯著減少(均P0.05),轉(zhuǎn)染sh EZH2后Eca109及TE1細(xì)胞遷移的距離明顯減小(均P0.05)。結(jié)論:EZH2可增加靶基因啟動(dòng)子上組蛋白H3第27位賴氨酸的三甲基化,并增強(qiáng)ESCC細(xì)胞的遷移和侵襲能力。
[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 鹵4.10), (鹵4.04) vs (132.00 鹵4.00), (105.33 鹵3.51, P 0.05), respectively], and the migration distance was significantly increased [(63.6 鹵1.2)]. (62.5 鹵2.5) vs (23.0 鹵2.3), (21.2 鹵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.
【作者單位】： 河北醫(yī)科大學(xué)第四醫(yī)院科研中心;
【基金】：河北省科技支撐計(jì)劃資助項(xiàng)目(No.14277732D,No.152777184)~~
【分類號(hào)】：R735.1

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