| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2005: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Research Abstract |
This study was aimed at analyzing about gene expression regulation mechanism of cytochrome P4501A1(CYP1A1), the major drug metabolism oxidizing enzyme, in aspect of with the base level, IC50, or maximum induction, regarding the transcription activation of nuclear receptor aryl-hydrocarbon receptor(AhR).
1.Establishment of cell-lines for the modifier cloning. Human hepatoma HepG2 or mouse hepatoma Hepa1c1c7 was transfected with a EGFP reporter construct which was connected a mouse CYP1A1promoter, and then stable tranformants was selected. These cell-lines can monitor an AhR dependence transcription induction of CYP1A1 gene by fluorescence measurement.
2.Analysis of influence of the first intron on an AhR dependence transcription of human CYP1A1 gene by using IRES connected luciferase system. I generated an unique IRES construct vector, and by using it, then revealed that there were an effective sequence which can inhibit the induction of CYP1A1gene transcription by AhR-TCDD, in the internal genomic region of human CYP1A1 gene, especially inside of the first intron.
3.A screening of genes which modulate benzopyrene resistance of the mouse hepatoma Hepa1c1c7 by using randomized hybrid ribozyme library. A randomized hybrid ribozyme library was introduced to the EGFP-reporter cell line established above(1). Among these transfected cell groups, a high group of EGFP expression strength by 3-methylchorantrene were separated with a cell-sorter machine and then collected the library from these cells.
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