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目的:观察转染RECK基因对人成釉细胞瘤(ameloblastoma,AM)细胞株hTERT+-AM的MMPs表达及细胞侵袭力的影响。方法:利用RECK基因慢病毒载体Lenti-RECK-eGFP/puro转染hTERT+-AM细胞株,Puro筛选和镜下挑单克隆纯化细胞。CCK-8检测细胞活性,Transwell实验检测细胞迁移、侵袭能力,qRT-PCR检测细胞中RECK的mRNA的表达情况。Western蛋白印迹检测细胞中RECK、MMP-2、MMP-9的表达情况。采用SPSS 13.0软件包对数据进行统计学分析。结果:转染RECK基因后,hTERT+-AM细胞中RECK的mRNA和蛋白的表达均显著增高(P均<0.01),细胞活性无显著变化(P均>0.05),细胞迁移、侵袭能力均显著下降(P均<0.01),MMP-2、MMP-9蛋白表达均显著下降(P均<0.05)。结论:RECK基因参与人成釉细胞瘤局部侵袭的调节,过表达RECK基因后,MMP-2、MMP-9下调,抑制AM细胞迁移和侵袭,RECK有望成为人成釉细胞瘤侵袭防治的新靶点。
Objective: To investigate the effect of transfected RECK gene on MMPs expression and cell invasiveness in human ameloblastoma (AM) cell line hTERT + -AM. Methods: Lenti-RECK-eGFP / puro was transfected into hTERT + -AM cell line by RECK lentivirus vector. The cell viability was detected by CCK-8, the migration and invasion ability of cells were detected by Transwell assay, and the mRNA expression of RECK was detected by qRT-PCR. Western blot was used to detect the expression of RECK, MMP-2 and MMP-9 in the cells. Data were statistically analyzed using SPSS 13.0 software package. Results: The RECK mRNA and protein expression in hTERT + -AM cells were significantly increased after transfected with RECK gene (all P <0.01), with no significant changes in cell activity (all P> 0.05). The migration and invasion ability of RECK cells were significantly decreased (All P <0.01), and the protein expressions of MMP-2 and MMP-9 decreased significantly (all P <0.05). CONCLUSIONS: RECK gene is involved in the regulation of local invasion of human ameloblastoma. After over-expression of RECK gene, MMP-2 and MMP-9 are down-regulated and the migration and invasion of AM cells are inhibited. RECK is expected to become a new target for prevention and treatment of human ameloblastoma point.