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 and Activity in Nasopharyngeal Carcinoma Cells

作　　者：王柏琦[1,2] 陈艳华[1] 蒋丽琴[1] 程爱兰[2] 

机构地区：[1]南华大学附属第二医院肿瘤内科,湖南衡阳421001 [2]南华大学肿瘤研究所

出　　处：《中南医学科学杂志》2014年第2期116-119,138,共5页Medical Science Journal of Central South China

基　　金：国家自然科学基金(81372894)

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、30μmol/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。结果 CNE-1细胞未刺激时RECK表达水平较低,而经1--30μmol/L姜黄素处理后,能显著增强RECK蛋白和mRNA的表达。高效液相色谱-电喷雾质谱结果显示,30μmol/L姜黄素处理后,RECK启动子、全基因组以及细胞核内甲基化水平分别降低为（69.04±10.62）%、（61.13±7.08）%、2.80±1.32。同时,姜黄素能显著降低CNE-1细胞中MMP-9蛋白和mRNA表达以及MMP-9的酶活性。结论姜黄素可能通过能上调RECK基因表达,降低细胞内甲基化水平,从而抑制MMP-9的表达与活性而发挥对CNE-1细胞生长抑制作用。Objective To 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. Methods Nasopharyngeal 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,re-spectively. Results The expression level of RECK was very low in unstimulated cells,and 1~30μmol/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. Conclusion Cucrcumin increase RECK gene expression and down-regulate its methylation,and then attenuate MMP-9 to inhibit CNE-1 cells growth.

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 

分 类 号：R739.62[医药卫生—肿瘤]