EZH2基因对人食管癌细胞增殖的影响

目的:探讨过表达或沉默果蝇Zeste基因增强子人类同源物2(enhancer of zeste homolog 2,EZH2)基因对食管癌细胞增殖的影响.方法:选用人食管癌细胞株ECA 109、TE1、KYSE30、KYSE 170作为研究对象,采用实时荧光定量PCR(qPCR)、Western blotting法分别检测食管癌细胞EZH2mRNA和蛋白的表达水平,然后采用qPCR检测过表达和沉默EZH2基因后对4株食管癌细胞EZH2mRNA的表达变化;CCK-8增殖实验、克隆形成实验检测过表达和沉默EZH2基因及EZH2抑制剂DZNep(3-deazaneplanocin A)处理对食管癌细胞增殖能力和克隆形成率的影响.结果:食管癌ECA 109、TE1细胞中EZH2 mRNA和蛋白水平明显高于KYSE30、KYSE 170细胞(P＜0.05).食管癌TE1、ECA 109细胞转染EZH2-ShRNA后EZH2表达水平下调(均P＜ 0.05)、细胞增殖能力降低(1.07±±0.08 vs 1.59±±0.09,P＜0.05;0.88±0.08 vs 1.05±0.11,P＜0.05)、克隆形成数下调[(200.00±11.43) vs (480.00±±13.10)个,P＜0.05;(88.00±±8.16) vs (220.00±±±14.69)个,P＜0.05].KYSE30、KYSE 170细胞转染EZH2过表达质粒后EZH2表达水平升高(均P＜0.05)、细胞增殖能力显著增强(1.06±0.07 vs 0.76±0.06,P＜0.05;3.36±±0.30 vs 1.50±0.08,P＜0.05)、克隆形成数显著升高[(45.00±3.27) vs (18.00±±1.63)个,P＜0.05;(65.00±4.08) vs (23.00±±2.45)个,P＜0.05];DZNep处理后,ECA109和TE1细胞增殖能力降低(均P＜0.05)、克隆形成数下降(均P＜0.05).结论:EZH2基因能有效促进食管癌细胞的增殖和克隆形成能力,为深入研究EZH2作为食管癌治疗的新靶点提供了实验研究基础.

Effect of EZH2 gene on proliferation of human esophageal cancer cells

Objective:To investigate the effect of EZH2 (enhancer of zeste homolog 2) overexpression or knockdown on the proliferation of esophageal cancer cells.Methods:Human esophageal cancer cell lines ECA 109,TE1,KYSE30 and KYSE170 were selected as the research objects.The expression ofEZH2 mRNA and protein in esophageal carcinoma cells were detected by Real-time fluorescence quantitative PCR (qPCR) and Western blotting,respectively.The mRNA expression of four esophageal cancer cells after overexpression or knockdown of EZH2 was detected by qPCR.The effects ofEZH2 overexpression or knockdown as well as EZH2 inhibitor DZNep (3-deazaneplanocin A) on the proliferation and clone growth rate of esophageal cancer cells were observed by CCK-8 proliferation assay and clone formation assay.Results:The expression of EZH2 mRNA and protein in ECA109 and TE1 cells was significantly higher than that in KYSE30 and KYSE170 cells (P＜0.05).The expression of EZH2 in esophageal cancer TE1 and ECA109 cells was down-regulated after transfection with EZH2-ShRNA,and the cell proliferation was decreased (1.07±0.08 vs 1.59±0.09,P＜0.05;1.05±0.11 vs 0.88±0.08,P＜0.05),and the clone formation was also down-regulated (200.00±11.43 vs 480.00±13.10,P＜0.05;88.00±8.16 vs 220.00±14.69,P＜0.05).The expression of EZH2 was increased in esophageal cancer KYSE30 and KYSE170 cells after transfection of EZH2 overexpression plasmid,and the proliferation ability (1.06±0.07 vs 0.76±0.06,P＜0.05;3.36±0.30 vs 1.50±0.08,P＜0.05)and clone formation (45.00±-3.27 vs 18.00±1.63,P＜0.05;65.00±4.08 vs 23.00±2.45,P＜0.05) were significantly increased.After DZNep treatment,proliferation (all P＜0.05) and clone formation (all P＜0.05) in ECA109 and TE1 cells were decreased.Conclusion:EZH2 gene can effectively promote the proliferation and cloning ability of esophageal cancer cells,which provides a basis for further study on the mechanism of EZH2 as a target in the treatment of esophageal cancer.