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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(P<0.05)。食管癌TE1、ECA109细胞转染EZH2-Sh RNA后EZH2表达水平下调(均P<0.05)、细胞增殖能力降低(1.07±0.08 vs1.59±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、KYSE170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。
Objective: To investigate the effect of enhancer of zeste homolog 2 (EZH2) gene on the proliferation of esophageal carcinoma cells. Methods: The human esophageal cancer cell lines ECA109, TE1, KYSE30 and KYSE170 were selected as the research objects. The expression of EZH2 mRNA and protein in esophageal cancer cells was detected by qPCR and Western blotting respectively. Then qPCR The expression of EZH2 m RNA in the four esophageal cancer cells was detected by overexpression and silencing of EZH2 gene. The proliferation of EKH2 m RNA was detected by CCK-8 proliferation assay and clone formation assay. The overexpression and silencing of EZH2 gene and DZNep inhibitor 3-deazaneplanocin A Effects of esophageal cancer cell proliferation and colony formation rate. Results: The mRNA and protein levels of EZH2 in esophageal cancer ECA109 and TE1 cells were significantly higher than those in KYSE30 and KYSE170 cells (P <0.05). The expression of EZH2 in esophageal cancer TE1 and ECA109 cells was down-regulated after EZH2-Sh RNA transfection (all P <0.05), and the cell proliferation was decreased (1.07 ± 0.08 vs 1.59 ± 0.09, P <0.05; 0.88 ± 0.08 vs 1.05 ± 0.11, P <0.05). The number of colony formation was 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]. EZH2 expression was increased in KYSE30 and KYSE170 cells transfected with EZH2 overexpression plasmid (all P <0.05), and cell proliferation was significantly increased (P <0.05; 3.36 ± 0.30 vs. 1.50 ± 0.08, P <0.05) (P <0.05). The number of colony formation was significantly higher than that of the control group (P <0.05), and the number of colony formation was significantly increased [(45.00 ± 3.27) vs (18.00 ± 1.63) vs The proliferation of ECA109 and TE1 cells decreased (all P <0.05), and the number of colony formation decreased (all P <0.05). CONCLUSION: EZH2 gene can effectively promote the proliferation and clonality of esophageal cancer cells, providing experimental basis for further study of EZH2 as a new target for the treatment of esophageal cancer.