摘要
目的：通过研究基质金属蛋白酶9(matrix metalloproteinase 9，MMP9)启动子片段的活性以及阿司匹林(aspirin)对启动子活性的调控，了解阿司匹林在前列腺癌的防治方面的机制。方法：DU145细胞体外培养，分别在24、48、72h采用MTF法检测不同浓度的阿司匹林对Du145的毒性。构建含人MMP-9基因5侧翼序列-1．28、-0．67、0．54、0．52kb与报告基因氯霉素乙酰基转移酶(CAT)的重组体，用Lipofetamine 2000转染试剂瞬时转染DU145细胞，CAT-ELISA测定各重组体的CAT值，及在乙酸肉豆蔻佛波酯(PMA)和／或阿司匹林作用下各重组体的CAT值。结果：低浓度的阿司匹林(≤1mmol·L^(-1))对DU145细胞毒性无明显差别。在无乙酸肉豆蔻佛波酯刺激的作用下，pCAT 1．28和pCAT 0．65的CAT表达量分别为对照组的2倍和1．6倍，而在乙酸肉豆蔻佛波酯的刺激下，pCAT 1．28、pCAT0．65和pCAT 0．54的CAT表达量分别为对照组的2．5、2．2和1．3倍。阿司匹林对pCAT 1．28、pCAT 0．65活性具有抑制作用且这种抑制作用可以被乙酸肉豆蔻佛波酯所抵消。结论：MMP9启动子的活性区域主要集中于-1285～-523之间的区域；阿司匹林可以在转录水平上下调MMP-9的表达，从而可能抑制前列腺癌的发生发展。
AIM： To understand the role of aspirin in the prevention and cure of prostatic carcinoma, and the effects of Aspirin on the promoter activities of human MMP-9 gene were studied. METHODS： DU145 cells were cultured and subcultured. Different concentrations of Aspirin （10, 5, 2.5, 1, 0.5, 0.1 and 0.2mmol·L^-1） were added into the plates to co-incubated with the cells. MTF method was used to detect cytotoxicity of Aspirin after 24, 48 and 72 hours. Cells were transient transfected with pCAT 1.28, pCAT 0.65, pCAT 0.54 and pCAT 0.52 containing - 1.28 kb, 0.65 kb, 0.54 kb, 0.52 kb of 5 flanked sequence of human MMP-9 gene, fused to CAT reporter gene by CAT-ELISA. RESULTS： Aspirin （ ≤ 1 mmol·L^-1） had no cytotoxicity of DU145 cells （ P 〉 0.05）. CAT expression of pCAT 1.28 and pCAT 0.65 are 2 and 1.6 times as high as the control group＇s. CAT expression of pCAT 1.28, pCAT 0.65 and pCAT 0.54 were 2.5, 2.2 and 1.3 times as high as the control group＇s. Aspirin inhibited the promoter activities of pCAT 1.28, pCAT 0.65. And the inhibition of Aspirin was reversed by PMA. CONGCLUSION： Aspirin can inhibit the expression of human MMP-9 gene in the transcriptional level and can inhibit the formation and development of prostatic carcinoma.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2005年第10期1190-1193,共4页
Chinese Journal of Clinical Pharmacology and Therapeutics
