Ery.43 a recent study of the n Be Tie-2 with suspensions of drugs has shown that decreases in vivo transscleral retinal delivery with a Erh Increase the corticosteroid transport by solubility.44 SCRPE will adversely by drugs Chtigt binding to melanin in the RPE layer choro and the tight junctions in the EPR layer.28 Grand in vitro and in vivo delivery of CSA indicates that they are able, it is to overcome both. The cytotoxicity t shown by CSA in human retinal pigmented epithelial cells that comprises the CSA no significant toxicity t in which M is 0.1 to 1 mM concentration range. Zus Tzlich for training pro-drug to reduce the cytotoxic effect. W While the CSA is not cytotoxic for 19 to 1 mM, celecoxib, in a previous study showed anything similar ARPE cell death by 50% to 49 M.45 Thus, the prodrug is not cytotoxic and k nnte For clinical application. improved therapeutic results, the closing will contribute to patient Lich Lebensqualit t. It is therefore the primary objective of this study to the M Possibility of clone 880 8 promoter, which are used for such a strategy check k Nnten. Materials and methods Cells and bacteria, a number of human prostate cancer LNCaP, was purchased by Bank Health Sciences Research Source.
The cells were grown and f in RPMI 1640 medium with 10% Fetal K Calf serum, antibiotics erg and optionally maintained at 37 1C in a humidified atmosphere of 5% CO 2 re Complements. The Escherichia coli strain DH5a was used for DNA manipulations experiments. The cells of E. coli were grown in LB medium at 37 1C. All compositions medium were purchased from BD Diagnostics. DNA manipulation experiments with E. coli were cozy the method of Sambrook et constructions Russell.16 clone clone 880 8880 8-promoter is a promoter to radiation artificially constructed, the expression of the luciferase gene is connected, obtained by 10 carried out as described ht, 4 times 12 h after exposure compared to R at 10 Gy were ntgenstrahlen concrete methods to construct the promoter told elsewhere.6 In short, a promoter probe vector, pGL3 DUTATA was described by cloning a reaction formed in each no polymerase amplified DNA fragment containing the signal of the box They TATA promoter in human H Controlled moxygenase gene upstream Rts of the I gene for luciferase pGL3 On.
Equimolar amount of the cis-elements for the synthesis of transcription factors that are sensitive to radiation confinement in prostate cancer cells, Lich NF kB, AP 1, 1 October, p53 and Nrf 2 additional Tzlich to 1/100 molar amounts of fragments of DNA sequences, Hind III recognition site restriction enzymes KpnI and were Feeder Ligated llig. After the ligated fragments were digested Rocuronium with HindIII and KpnI, the resulting sequences were then into the KpnI and HindIII sites of pGL3 Tata, the cloned immediately before the signal bo They TATA, the construction of 28 plasmids form a promoter library. These plasmids were then transduced in LNCaP cells and a stimulation of the radiation. We have the most reactive are 6.7-fold improvement in Luciferaseaktivit t 48 h after 10 Gy R-Rays, designated clone 880-promoter. The sequences Age analysis showed that was 386 nucleotides long and consisted of two cis-elements NF kB, 1 October cis-element, four cis-elements, p53 and three cis-elements NRF 2, but no element AP cis-1. In step n, Mutat HIGHEST Feeder Llige.