地西他滨诱导miR-200c/141表达抑制肾癌细胞侵袭迁移

Decitabine Inhibit Invasion and Migration Ability of Renal Cell Carcinoma Cells by Up-regulating MiR-200c/141

关键词:  地西他滨  miR-200c  miR-141  肾细胞癌  侵袭迁移

Key words:  decitabine  miR-200c  miR-141  renal cell carcinoma  invasion and migration

摘要:

目的 探讨DNA甲基转移酶抑制剂地西他滨对人肾细胞癌786-O和769-P细胞侵袭迁移能力的影响，并进行相关机制的研究。方法 实时荧光定量PCR法检测肾癌及其癌旁组织中miR-200c/141的表达量及2.5 μmol·L^-1地西他滨对肾癌细胞系miR-200c/141的诱导作用；Transwell试验、划痕试验分别检测地西他滨、miR-200c/141对细胞侵袭迁移的影响；实时荧光定量PCR法、蛋白印迹法检测786-O和769-P细胞系钙黏蛋白表达量。结果 与正常肾癌旁组织相比，miR-200c/141在肾癌组织中的表达量显著下调。2.5 μmol·L^-1地西他滨能诱导786-O和769-P细胞系中miR-200c/141的表达增加。地西他滨、miR-200c/141均能抑制细胞的侵袭迁移能力。786-O和769-P细胞经地西他滨处理后，与上皮间充质转化相关的钙黏蛋白表达水平显著上调。结论 地西他滨可以通过上调miR-200c/141的表达水平减弱肾癌细胞系的侵袭迁移能力。

Abstract:

OBJECTIVE To investigate the effect and mechanism of DNA methyltransferase inhibitor decitabine on the invasion and migration ability of renal cancer cells 786-O and 769-P. METHODS The miR-200c/141 expression levels in renal cell carcinoma and its adjacent tissues were detected by RT-qPCR. MiR-200c/141 were induced by 2.5 μmol·L^-1 decitabine and detected by RT-qPCR. Transwell assay and scratch assay were used to detect the effect of decitabine, miR-200c/141 on cell invasion and migration respectively. The mRNA and protein levels of E-cadherin were detected by RT-qPCR and western blot. RESULTS Compared with adjacent tissues, miR-200c/141 were significantly down regulated in renal cell carcinoma. The expression of miR-200c/141 could be induced in the 786-O and 769-P cells after treated with 2.5 μmol·L^-1 decitabine. Scratch assay and Transwell assay results showed that decitabine, miR-200c/141 could inhibit cell migration and invasion. The expression of E-cadherin was upregulated in 786-O and 769-P cells after treated with decitabine, which associated with epithelial mesenchymal transition. CONCLUSION Decitabine can weaken the migration and invasion ability of 786-O and 769-P cells by up-regulating the expression of miR-200c/141.