王柏琦,,陈艳华,蒋丽琴,程爱兰,.姜黄素对鼻咽癌细胞RECK基因甲基化以及MMP-9表达与活性的影响.[J].中南医学科学杂志.,2014,42(2):116-119. |
姜黄素对鼻咽癌细胞RECK基因甲基化以及MMP-9表达与活性的影响 |
Effect of Curcumin on RECK Methylation and MMP-9 Expression andActivity in Nasopharyngeal Carcinoma Cells |
投稿时间:2013-11-17 |
DOI: |
中文关键词: 鼻咽癌 回复引导半胱氨酸丰富蛋白含kazal基元 甲基化 姜黄素 |
英文关键词:nasopharyngeal carcinoma reversion-inducing-cysteine-rich protein with kazal motifs methylation |
基金项目:国家自然科学基金(81372894). |
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中文摘要: |
目的探讨姜黄素对鼻咽癌细胞回复引导半胱氨酸丰富蛋白含kazal基元(RECK)基因甲基化以及基质金属蛋白酶9(MMP-9)表达的影响。方法体外培养鼻咽癌细胞系CNE-1,用1、10、30 μmol/L姜黄素处理后,采用Western blot和实时定量PCR分别检测RECK、MMP-9蛋白和mRNA表达;高效液相色谱-电喷雾质谱检测RECK甲基化;MTT法检测CNE-1细胞的增殖。同时采用明胶酶谱实验观察姜黄素处理前后MMP-9酶活性变化。 |
英文摘要: |
ObjectiveTo investigate the effect of curcumin on reversion-inducing-cysteine-rich protein with kazal motifs (RECK) gene methylation,and expression of matrix metalloproteinase-9 (MMP-9) in nasopharyngeal carcinoma cells.MethodsNasopharyngeal carcinoma cell line CNE-1 was cultured in vitro,and stimulated by 1,10 and 30 μmol/L curcumin for 48h,expression of RECK and MMP-9 were detected by Western blot and real-time PCR,respectively.RECK methylation was determined by HPLC chromatographic and mass spectrometric methods.Cell proliferation was assessed by MTT assay.Expression and the enzymic activity of MMP-9 were detected by Western blot and Gelatin zymography assay,respectively.ResultsThe expression level of RECK was very low in unstimulated cells,and 1~30 μmol/L could decrease the protein and mRNA level.HPLC chromatographic and mass spectrometric analysis demonstrated that 30 μmol/L could decrease level of the promoter methylation,global DNA methylation and the methylation activity of the nuclear extract to (69.04%±10.62)%、(61.13±7.08)%、2.80±1.32,respectively,as compared to untreated cells.In addition,curcumin could also decrease the protein and mRNA level of MMP-9,and enzymic activity.ConclusionCucrcumin increase RECK gene expression and down-regulate its methylation,and then attenuate MMP-9 to inhibit CNE-1 cells growth. |
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