EZH2基因对人食管癌细胞增殖的影响
发布时间：2018-11-04 16:18
【摘要】：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ECA109、TE1、KYSE30、KYSE170作为研究对象,采用实时荧光定量PCR(q PCR)、Western blotting法分别检测食管癌细胞EZH2 m RNA和蛋白的表达水平,然后采用q PCR检测过表达和沉默EZH2基因后对4株食管癌细胞EZH2 m RNA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ECA109、TE1细胞中EZH2 m RNA和蛋白水平明显高于KYSE30、KYSE170细胞(P0.05)。食管癌TE1、ECA109细胞转染EZH2-Sh RNA后EZH2表达水平下调(均P0.05)、细胞增殖能力降低(1.07±0.08 vs1.59±0.09,P0.05;0.88±0.08 vs 1.05±0.11,P0.05)、克隆形成数下调[(200.00±11.43)vs(480.00±13.10)个,P0.05;(88.00±8.16)vs(220.00±14.69)个,P0.05]。KYSE30、KYSE170细胞转染EZH2过表达质粒后EZH2表达水平升高(均P0.05)、细胞增殖能力显著增强(1.06±0.07 vs 0.76±0.06,P0.05;3.36±0.30 vs 1.50±0.08,P0.05)、克隆形成数显著升高[(45.00±3.27)vs(18.00±1.63)个,P0.05;(65.00±4.08)vs(23.00±2.45)个,P0.05];DZNep处理后,ECA109和TE1细胞增殖能力降低(均P0.05)、克隆形成数下降(均P0.05)。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。
[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 卤0.08 vs1.59 卤0.09 vs1.59 卤0.05) after transfection of EZH2-Sh RNA. 0. 88 卤0. 08 vs 1. 05 卤0. 11 vs. The number of clones was down-regulated [(200.00 卤11. 43) vs (480.00 卤13. 10), P 0. 05; (88.00 卤8.16) vs (220.00 卤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 卤0.07 vs 0.76 卤0.06P0.05; 3. 36 卤0. 30 vs 1. 50 卤0. 08 vs, significantly increased the number of clone formation [(45. 00 卤3. 27) vs (18. 00 卤1. 63), P 0. 05; (65. 00 卤4. 08) vs (23. 00 卤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.
【作者单位】： 河北医科大学第四医院肿瘤研究所免疫学实验室;
【基金】：河北省科技支撑计划资助项目(No.152777184)~~
【分类号】：R735.1
[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 卤0.08 vs1.59 卤0.09 vs1.59 卤0.05) after transfection of EZH2-Sh RNA. 0. 88 卤0. 08 vs 1. 05 卤0. 11 vs. The number of clones was down-regulated [(200.00 卤11. 43) vs (480.00 卤13. 10), P 0. 05; (88.00 卤8.16) vs (220.00 卤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 卤0.07 vs 0.76 卤0.06P0.05; 3. 36 卤0. 30 vs 1. 50 卤0. 08 vs, significantly increased the number of clone formation [(45. 00 卤3. 27) vs (18. 00 卤1. 63), P 0. 05; (65. 00 卤4. 08) vs (23. 00 卤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.
【作者单位】： 河北医科大学第四医院肿瘤研究所免疫学实验室;
【基金】：河北省科技支撑计划资助项目(No.152777184)~~
【分类号】：R735.1
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