NORE1基因在肾透明细胞癌细胞迁移侵袭和血管生成中的作用及相关机制的研究

摘要

目的 探讨NORE1基因过表达对肾透明细胞癌细胞迁移侵袭及血管生成的影响及其分子作用机制.方法 利用NORE1过表达质粒和对照组质粒,分别转染人肾透明细胞癌786-O和ACHN细胞,Western blot检测细胞转染效率,Transwell细胞迁移实验和侵袭实验检测细胞迁移和侵袭的能力,体外血管生成实验观察其血管生成能力,Western blot检测NORE1基因过表达后肾癌细胞中基质金属蛋白酶(MMPs)的蛋白表达水平.结果 转染NORE1过表达质粒后,肾透明细胞癌786-O、ACHN细胞中NORE1表达量增加.NORE1基因过表达后肾透明细胞癌细胞迁移及侵袭能力减弱,血管生成能力减弱,差异均具有统计学意义(P＜0.001).NORE1过表达组肾透明细胞癌细胞中MMP 9表达较对照组显著下降,而两组间MMP-2表达水平无显著性差异.结论 肾透明细胞癌细胞中NORE1基因可以通过调节MMP-9的表达影响细胞的迁移侵袭及血管生成.%Objective To investigate the effects of novel Ras effector 1 (NORE1) overexpression on the migration,invasion and angiogenesis of human clear cell renal cell carcinoma (ccRCC) cell lines 786-O and ACHN,and to explore the related molecular mechanisms.Methods NORE1 overexpression plasmid and negative control were transiently transfected into 786-O and ACHN cell lines using LipofectamineTM 2000.Western blotting was performed to measure the expression of NORE1 and MMPs.The cellular capabilities of migration,invasion and angiogenesis were examined by Transwell assay and vitro angiogenesis experiment respectively.Results The NORE1 overexpression plasmid could remarkably enhance the quantities of NORE1 in both 786-O and ACHN cell lines.The cellular capabilities of migration,invasion and angiogenesis were significantly decreased (P＜0.001).Specific decrease of MMP-9 expression was noticed in both 786-O and ACHN cell lines with the overexpression of NORE1;meanwhile,the MMP-2 expression remained the same.Conclusion NORE1 in ccRCC cells can affect cell migration,invasion and angiogenesis by regulating the expression of MMP-9.