The Effects Of Aspirin On The Promoter Activities Of Human MMP-9 Gene In DU145 Cells

AIM: To infer the mechanism of Aspirin in the prevention and cure of Prostatic Carcinoma, the effects of Aspirin on the promoter activities of human MMP-9 gene were studied.METHODS: 1. Construct the expressive vector of pCAT1.28, pCAT0.65, pCAT0.54 and pCAT0.52 containing CAT and -1.28kb, -0.65kb, -0.54kb, -0.52kb of 5 flanked sequence of human MMP-9 gene. 2. DU145 cells were cultured and subcultured. Different concentrations of Aspirin (10 mmol-L-1, 5 mmol-L-1, 2.5 mmol-L-1, 1 mmol-L-1, 0.5 mmol-L-1, 0.1 mmol-L-1, 0.02 mmol-L-1 ) were added into the plates to co-incubated with the cells. MTT method was used to detect cytotoxicity of Aspirin after 24, 48 and 72 hours. 3. The cells were grouped for control, PMA, Aspirin and PMA+Aspirin. DU145 cells were transient transfected with expressive vector. Then determined results of CAT through CAT-Elisa.RESULTS: 1. Succeed to construct the expressive vector of pCAT1.28, pCAT0.65, pCAT0.54 , pCAT0.52. 2. Aspirin (0.05). 3. When not stimulated by PMA, CAT expression of pCAT1.28 and pCAT0.65 were 2 and 1.6 times as high as CAT expression of the control group. When, CAT expression of pCAT1.28, pCAT0.65 and pCAT0.54 were 2.5, 2.2 and 1.3 times as high as CAT expression of the control group. Even stimulated by PMA, CAT expression of pCAT0.52 weren't higher. Aspirin can inhibit the promoter activities of pCAT1.28, pCAT0.65. And the inhibition of Aspirin can be partly or totally reversed by PMA.CONGCLUSIONS: 1. Expressive vectors of pCAT1.28 , pCAT0.65 ,pCATO.54 , pCATO.52 almost include all of active sites of MMP-9 promoter. 2. These data indicate that Aspirin can inhibit the expression of human MMP-9 gene in the transcriptional level and may inhibit the formation and development of prostate carcinoma.