Background and objective: The anti-tumor activity of dexamethasone derivatives (9-fluoro-16alpha-methyl-11,17-dihydroxy-3-oxo-1,4-androsladiene-17beta-carboxylic acid) is superior to that of dexamethasone. This study was to screen the proteins interacting with dexamethasone derivates, thus to explore the anti-tumor mechanism of dexamethasone derivates in vivo.
Methods: The bait plasmid pGBKT7-GRalpha-LBD was constructed. Screening of the target proteins interacting with dexamethasone derivatives was performed by yeast three-hybrid technique using human K562 cell cDNA library.
Results: The bait plasmid was successfully constructed. It produced a 31 ku bait protein with no toxicity, leakage and self-activation. Thirty-seven positive clones which interacted with dexamethasone derivatives were obtained from human K562 cell cDNA library, 20 of which were identified by re-transforming into yeast AH109 cells.
Conclusion: Twenty positive clones interacting with dexamethasone derivates are identified in vivo.