5-氮杂-2′-脱氧胞苷对腺样囊性癌细胞 RECK 基因表达及细胞侵袭力的影响

目的：探讨甲基化转移酶抑制剂5-氮杂-2′-脱氧胞苷（5-aza-2′deoxycytidine，5-aza-dC）对腺样囊性癌细胞中 RECK基因表达及肿瘤侵袭力的影响。方法：采用甲基化特异性 PCR、实时定量 PCR、Western 印记检测腺样囊性癌细胞中 RECK 基因的甲基化状态及 RECK 基因 mRNA 和蛋白的表达，transwell 体外侵袭实验检测细胞侵袭能力。结果：RECK 基因甲基化条带仅在 ACC-M细胞中发现，经5-aza-dC 处理后 ACC-M细胞中 RECK 基因甲基化得到逆转，RECK 基因 mRNA 及蛋白的表达显著提升。ACC-M细胞的侵袭力明显降低。结论：5-aza-dC 可逆转腺样囊性癌 ACC-M细胞中 RECK 基因的高甲基化状态并恢复 RECK 基因 mRNA 及蛋白的表达，抑制 ACC-M细胞侵袭力。

The effects of 5-aza-2-deoxycytidine on RECK gene expression and invasion of salivary adenoid cystic car-cinoma cells

Objective:To investigate the effects of 5-aza-2′deoxycytidine(5-aza-dC),a DNA methyltransferase (DNMT)inhibitor, on the methylation status of the RECK gene and the invasion of salivary adenoid cystic carcinoma cell lines.Methods:Methylation-specific PCR,Western blot analysis and quantitative real-time PCR were used to investigate the methylation status of RECK gene and the expression of RECK mRNA and protein in SACC cell lines.The invasive ability of SACC cells was examined by transwell assay. Results:Promoter methylation was only found in ACC-Mcell line and not in ACC-2 cell line.Treatment of ACC-Mcells with 5-aza-dC partially reversed the hypermethylation status of the RECK gene and significantly enhanced the expression level of mRNA and pro-tein of RECK,suppressed ACC-Mcell invasive ability.Conclusion:5-aza-dC can inhibit ACC-Mcell invasion by reversal of hyperm-ethylation status of RECK gene.