[Establishment of reporter gene-based cell screening model for discovering new chemopreventive agents]

Area of Science:

• Biochemistry
• Molecular Biology
• Pharmacology

Background:

• Chemopreventive agents hold promise for cancer prevention.
• Developing efficient drug screening models is crucial for discovering new chemopreventive compounds.
• Reporter gene assays provide a sensitive method for monitoring cellular responses.

Purpose of the Study:

• To establish a novel cell-based drug screening model for identifying chemopreventive agents.
• To utilize an antioxidative response element (ARE) linked to a reporter gene (GFP) for monitoring chemopreventive activity.
• To validate the model's efficacy using known chemopreventive compounds.

Main Methods:

• Synthesized and cloned ARE DNA binding sequences into a GFP expression vector.
• Stably transfected the construct into HepG2 cells to create a reporter cell line.
• Assessed the dose-dependent effects of known chemopreventive agents (resveratrol, protocatechuic aldehyde, ursolic acid, oleanolic acid) on GFP expression.

Main Results:

• The antioxidative response element (ARE) successfully regulated GFP induction in a dose-dependent manner within a specific concentration range.
• Resveratrol significantly increased the level of GFP expression, indicating its chemopreventive potential.
• The cell model demonstrated sensitivity to known chemopreventive agents.

Conclusions:

• The established reporter gene-based cell model is effective for screening chemopreventive agents.
• This method facilitates the identification of novel chemopreventive compounds, particularly from traditional Chinese medicine sources.
• Measurement of GFP luminescence in microtiter plates provides a quantitative readout for drug screening.