Recombination method and application of luciferase reporter gene with aquaporin 1 (AQP1) promoter to drug screening
A technology of aquaporin and reporter gene, which is applied in the field of constructing AQP1 promoter salivary gland cell model and drug screening, and can solve the problems that there are no therapeutic drugs.
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[0068] Example 2: Drug screening
[0069] 1. Cultivation of cell lines
[0070] Adenoid cystic cancer cells were cultured in DMEM, supplemented with 10% fetal bovine serum, 100U / mL penicillin and 100ug / mL streptomycin. The cells were cultured in a 5% incubator at 37°C.
[0071] 2. Prove the effectiveness of the drug screening model
[0072] Scholars at home and abroad have confirmed that estradiol (E 2 ) Plays an important role in the gene expression of AQP1. The present invention uses AQP1 promoter luciferase reporter gene recombinant plasmid cell model to confirm that E 2 It can activate the AQP1 promoter in a dose-dependent manner [ Figure 8 】At the same time, we also confirmed that estradiol can promote the expression of AQP1 protein【 Picture 9 ], which proves that the cell model can be used for drug screening simply and sensitively.
[0073] 3. Application of drug screening
[0074] First take 2×10 5 ACC-M cells are plated in a 48-well plate to make the cells reach a confluence r...
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