【摘要】：目的:探討過表達(dá)或沉默果蠅Zeste基因增強(qiáng)子人類同源物2(enhancer of zeste homolog 2,EZH2)基因?qū)κ彻馨┘?xì)胞增殖的影響。方法:選用人食管癌細(xì)胞株ECA109、TE1、KYSE30、KYSE170作為研究對(duì)象,采用實(shí)時(shí)熒光定量PCR(q PCR)、Western blotting法分別檢測(cè)食管癌細(xì)胞EZH2 m RNA和蛋白的表達(dá)水平,然后采用q PCR檢測(cè)過表達(dá)和沉默EZH2基因后對(duì)4株食管癌細(xì)胞EZH2 m RNA的表達(dá)變化;CCK-8增殖實(shí)驗(yàn)、克隆形成實(shí)驗(yàn)檢測(cè)過表達(dá)和沉默EZH2基因及EZH2抑制劑DZNep(3-deazaneplanocin A)處理對(duì)食管癌細(xì)胞增殖能力和克隆形成率的影響。結(jié)果:食管癌ECA109、TE1細(xì)胞中EZH2 m RNA和蛋白水平明顯高于KYSE30、KYSE170細(xì)胞(P0.05)。食管癌TE1、ECA109細(xì)胞轉(zhuǎn)染EZH2-Sh RNA后EZH2表達(dá)水平下調(diào)(均P0.05)、細(xì)胞增殖能力降低(1.07±0.08 vs1.59±0.09,P0.05;0.88±0.08 vs 1.05±0.11,P0.05)、克隆形成數(shù)下調(diào)[(200.00±11.43)vs(480.00±13.10)個(gè),P0.05;(88.00±8.16)vs(220.00±14.69)個(gè),P0.05]。KYSE30、KYSE170細(xì)胞轉(zhuǎn)染EZH2過表達(dá)質(zhì)粒后EZH2表達(dá)水平升高(均P0.05)、細(xì)胞增殖能力顯著增強(qiáng)(1.06±0.07 vs 0.76±0.06,P0.05;3.36±0.30 vs 1.50±0.08,P0.05)、克隆形成數(shù)顯著升高[(45.00±3.27)vs(18.00±1.63)個(gè),P0.05;(65.00±4.08)vs(23.00±2.45)個(gè),P0.05];DZNep處理后,ECA109和TE1細(xì)胞增殖能力降低(均P0.05)、克隆形成數(shù)下降(均P0.05)。結(jié)論:EZH2基因能有效促進(jìn)食管癌細(xì)胞的增殖和克隆形成能力,為深入研究EZH2作為食管癌治療的新靶點(diǎn)提供了實(shí)驗(yàn)研究基礎(chǔ)。
[Abstract]:Aim: to investigate the effect of overexpression or silencing of Zeste gene enhancer 2 (enhancer of zeste homolog 2EZH2) gene on the proliferation of esophageal cancer cells. Methods: the expression of EZH2 m RNA and protein in human esophageal carcinoma cell line ECA109,TE1,KYSE30,KYSE170 was detected by real-time fluorescence quantitative PCR (q PCR), Western blotting assay. Then Q PCR was used to detect the expression of EZH2 m RNA in four esophageal cancer cells after overexpression and silencing of EZH2 gene. The effects of overexpression and silencing of EZH2 gene and EZH2 inhibitor DZNep (3-deazaneplanocin A) on the proliferation and clone formation rate of esophageal carcinoma cells were detected by CCK-8 proliferation assay and clone formation assay. Results: the levels of EZH2 m RNA and protein in esophageal carcinoma ECA109,TE1 cells were significantly higher than those in KYSE30,KYSE170 cells (P 0.05). The expression of EZH2 was down-regulated (P0.05) and the proliferation ability of TE1,ECA109 cells was decreased (1.07 鹵0.08 vs1.59 鹵0.09 vs1.59 鹵0.05) after transfection of EZH2-Sh RNA. 0. 88 鹵0. 08 vs 1. 05 鹵0. 11 vs. The number of clones was down-regulated [(200.00 鹵11. 43) vs (480.00 鹵13. 10), P 0. 05; (88.00 鹵8.16) vs (220.00 鹵14.69, P0.05). The expression of EZH2 in KYSE30,KYSE170 cells increased after transfection of EZH2 overexpression plasmids (P0.05), and the proliferation ability of KYSE30,KYSE170 cells increased significantly (1.06 鹵0.07 vs 0.76 鹵0.06P0.05; 3. 36 鹵0. 30 vs 1. 50 鹵0. 08 vs, significantly increased the number of clone formation [(45. 00 鹵3. 27) vs (18. 00 鹵1. 63), P 0. 05; (65. 00 鹵4. 08) vs (23. 00 鹵2. 45), P0.05]; After DZNep treatment, the proliferation ability of ECA109 and TE1 cells decreased (P0.05), and the number of clone formation decreased (P0.05). Conclusion: EZH2 gene can effectively promote the proliferation and clone formation of esophageal cancer cells and provide experimental basis for further study of EZH2 as a new target of esophageal cancer therapy.
【作者單位】： 河北醫(yī)科大學(xué)第四醫(yī)院腫瘤研究所免疫學(xué)實(shí)驗(yàn)室;
【基金】：河北省科技支撐計(jì)劃資助項(xiàng)目(No.152777184)~~
【分類號(hào)】：R735.1

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