目的:探讨去甲基化试剂5-氮杂胞苷(5-azacytidine,5-azac)上调RUNX3基因表达后,食管癌Eca109细胞生物学行为的改变。方法:体外培养Eca109细胞,应用10、20和50μmol/L较低浓度的5-azac干预72h,实时定量PCR及蛋白质印迹法检测RUNX3基因的mRNA及蛋白表达水平改变;甲基化特异性PCR检测RUNX3基因的甲基化程度变化;细胞划痕、Transwell实验观察Eca109细胞迁移及侵袭能力的改变;裸鼠成瘤实验观察体内Eca109细胞移植瘤生长的变化。结果:经10、20和50μmol/L 5-azac干预72h后,RUNX3基因的mRNA表达水平分别增加2.18±0.23、4.57±0.26和6.90±0.36倍;RUNX3蛋白表达相对系数分别为0.196±0.004、0.278±0.013和0.396±0.025。Eca109细胞经5-azac处理后,部分RUNX3基因去甲基化。0、20和50μmol/L 5-azac干预72h后,RUNX3基因的去甲基化条带亮度值比值分别为0.125±0.003、1.791±0.112和2.987±0.236,P<0.001。0、10、20和50μmol/L 5-azac干预72h后,Eca109细胞运动至划痕处细胞数分别为715±20.1、398±19.3、302±16.4和124±13.8。相比于对照组,10、20和50μmol/L干预组迁移细胞数百分率分别为62%、46%和12%,侵袭细胞数百分率为60%、45%和8%。RUNX3基因表达上调后,Eca109细胞裸鼠体内移植瘤生长减慢,F=67.5,P<0.001;10、20及50μmol/L组肿瘤生长抑制率分别为25.6%、53.4%和79.8%。结论:较低浓度5-azac通过去甲基化途径上调人食管鳞癌Eca109细胞的RUNX3基因表达,并抑制Eca109细胞迁移、侵袭及体内增殖。 Objective: To investigate the biological behavior of esophageal cancer Eca109 cells after 5-azacytidine (5-azac) up-regulates RUNX3 gene expression. Methods: Eca109 cells were cultured in vitro. The mRNA and protein levels of RUNX3 gene were detected by real-time quantitative PCR and Western blotting after interference with 5-azac at 10, 20 and 50μmol / L for 72 hours. Methylation-specific PCR RUNX3 gene methylation changes; cell scratches, Transwell assay Eca109 cell migration and invasion ability changes; nude mice tumorigenicity experiment observed Eca109 cells in vivo tumor growth changes. Results: The mRNA expression levels of RUNX3 increased by 2.18 ± 0.23, 4.57 ± 0.26 and 6.90 ± 0.36 times after 10, 20 and 50μmol / L 5-azac treatment, respectively. The relative expression coefficients of RUNX3 protein were 0.196 ± 0.004 and 0.278 ± 0.013 and 0.396 ± 0.025. After Eca109 cells were treated with 5-azac, some RUNX3 genes were demethylated. After 72 hours of intervention with 0, 20 and 50μmol / L 5-azac, the brightness degeneration of RUNX3 gene was 0.125 ± 0.003,1.791 ± 0.112 and 2.987 ± 0.236, P <0.001.0,10,20 and After 72 hours of intervention with 50μmol / L 5-azac, the numbers of cells moving to scratch in Eca109 cells were 715 ± 20.1, 398 ± 19.3, 302 ± 16.4 and 124 ± 13.8, respectively. The percentage of migrating cells in the 10, 20 and 50 μmol / L intervention groups was 62%, 46% and 12%, respectively, and the percentage of invasive cells was 60%, 45% and 8%, respectively, compared to the control group. After RUNX3 gene expression was up-regulated, the growth of transplanted tumors in Eca109 cells in nude mice was slowed down (F = 67.5, P <0.001). The tumor growth inhibition rates were 25.6%, 53.4% ​​and 79.8% in 10, 20 and 50μmol / CONCLUSION: Lower concentration of 5-azac up-regulates the expression of RUNX3 gene in esophageal squamous cell carcinoma Eca109 cells through demethylation and inhibits the migration, invasion and proliferation of Eca109 cells.