UHRF1在胃癌侵袭和转移中的作用及其机制

Effect of Ubiquitin-like with PHD and Ring Finger Domain 1 on Invasion and Metastasis of Gastric Cancer and Related Mechanism

目的探讨甲基化调节因子UHRF1在胃癌转移过程中的作用及其调控机制。方法 RT-PCR和Western blot检测胃癌高低转移细胞系中UHRF1的表达,应用UHRF1的si RNA和质粒改变UHRF1的表达,Transwell及体内转移实验检测胃癌转移能力的变化。甲基化特异性PCR和Western blot检测UHRF1表达改变后p16^(INK4)和RB1的甲基化状态及蛋白表达。结果 UHRF1在高转移潜能的胃癌细胞系GC9811-P中的表达显著高于低转移潜能的细胞系GC9811。下调UHRF1的表达抑制胃癌细胞的体外和体内转移能力,同时减弱p16^(INK4)甲基化,增加p16^(INK4)蛋白表达。上调UHRF1的表达可见相反结果。结论 UHRF1可能通过调控p16^(INK4)和RB1的甲基化促进胃癌细胞的侵袭和转移,其有望成为胃癌转移预警和治疗的新靶点。

Objective To explore the role of ubiquitin-like with PHD and ring finger domain 1（UHRF1） in the invasion and metastasis of gastric cancer（GC） cells. Methods We examined UHRF1 expression in two pairs of highly metastatic and lowly metastatic GC cell lines by RT-PCR and Western blot. Western blot and in vivo metastasis assays were conducted to measure the migration and invasion abilities of GC cells after UHRF1 expression was altered by si RNA or vector. Then methylation specific PCR and Western blot were performed to detect the methylation status and the expression of p16INK4 A and RB1. Results UHRF1 expression was markedly higher in highly metastatic GC cells compared with their counterparts. Down-regulation of UHRF1 expression suppressed the invasion and metastasis of GC cells, and increased p16INK4 A and RB1 genes expression via promoter demethylation. Up-regulation of UHRF1 expression showed the opposite results. Conclusion UHRF1 could promote the invasion and metastasis of GC cells via regulating p16INK4 A and RB1 methylation. UHRF1 may become a new target for early warning and treatment of GC metastasis.