高浓度硝酸甘油对肿瘤细胞MMP-9/TIMP-1表达的调控

Regulation of MMP-9/TIMP-1 Expression in High Concentration GTN Treated Tumor Cells

利用基因芯片、RT-PCR技术和ELISA方法观察硝酸甘油(GTN)作为一氧化氮供体对人宫颈癌细胞(Hela)MMP-9/TIMP-1基因及蛋白表达的调控。通过基因芯片对肿瘤转移相关基因的表达进行筛选,应用RT-PCR技术对MMP-9/TIMP-1基因表达进行观察,ELISA方法测定细胞培养液中MMP-9/TIMP-1蛋白浓度并计算二者比值。结果显示,培养液GTN浓度为40μg/mL时,细胞中多个肿瘤相关基因表达改变;实验组与对照组比较,细胞MMP-9基因表达有大幅度下降,TIMP-1基因表达改变不明显;实验组细胞培养液MMP-9蛋白浓度也出现明显降低,TIMP-1无显著改变,MMP-9/TIMP-1比值变化显著。结论:硝酸甘油在一定浓度下能够调控肿瘤细胞的基因表达并改变MMP-9/TIMP-1的动态平衡。

To investigate the regulation of MMP-9/TIMP-1 expressions in glyceryl trinitrate（GTN） treated Hela cells.The Hela cells were treated with GTN that acted as the donor of nitric oxide.The microarray technique was performed to analyze gene expression profiles related tumor angiogenesis and metastasis.MMP-9 and TIMP-1 gene expressions in treated Hela cells were examined through RT-PCR.ELISA assay was applied to determine the protein concentrations of MMP-9 and TIMP-1 in culture medium and calculate the MMP-9/TIMP-1 ratio.The results showed that some gene expressions were changed in Hela cells treated with 40μg/ml GTN in culture medium.The expression of MMP-9 in GTN group was greatly lower than that in control group,but the expression of TIMP-1 was not obviously changed.MMP-9 concentration in culture medium reduced dramatically,but no changed for TIMP-1.The ratio of MMP-9/TIMP-1 was altered statistically.The results indicate that glyceryl trinitrate in certain concentration could regulate the gene expression in tumor cell and shift the production balance of MMP-9 and TIMP-1.