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目的:研究雷公藤内酯醇(triptolide,TL)抑制纤维肉瘤HT-1080细胞基质金属蛋白酶(matrix metalloproteinase,MMP-9)表达的机制。方法:用终浓度分别为6,12,18 nmol.L-1的TL处理HT-1080细胞72 h,对照组不加药物处理,甲基化特异性PCR检测TL对MMP-9基因启动子甲基化水平的影响,RT-PCR检测TL对基质金属蛋白酶组织抑制物(TIMP-1,-2,-3和-4 mRNA表达的影响。结果:与对照组比较,3个TL处理组中仅18 nmol.L-1处理组MMP-9基因甲基化指数明显上调(0.39±0.10 vs 0.61±0.10,P<0.05);所有TL处理组TIMP-1,-2,-3和-4的mRNA表达均无明显变化。结论:TL上调HT-1080细胞MMP-9基因甲基化水平,这可能是TL抑制肿瘤MMP-9表达的一种新机制。
Objective: To study the mechanism of triptolide (TL) inhibiting the expression of matrix metalloproteinase (MMP-9) in fibrosarcoma HT-1080 cells. METHODS: HT-1080 cells were treated with TL with a final concentration of 6, 12, and 18 nmol.L-1 for 72 h. The control group received no drug treatment. Methylation-specific PCR detected TL against MMP-9 gene promoter A. Effect of basal levels, RT-PCR detection of TL on tissue inhibitor of metalloproteinase (TIMP-1, -2, -3 and -4 mRNA expression. The results: compared with the control group, 3 TL treatment group only The methylation index of MMP-9 gene in 18 nmol.L-1 treatment group was significantly up-regulated (0.39±0.10 vs 0.61±0.10, P<0.05); mRNA of TIMP-1, -2, -3, and -4 in all TL treatment groups There was no significant change in expression.Conclusion: TL can up-regulate the methylation of MMP-9 gene in HT-1080 cells, which may be a new mechanism of TL inhibiting the expression of MMP-9 in tumors.