Screening of chemopreventive tea polyphenols against PAH genotoxicity in breast cancer cells by a XRE-luciferase reporter construct.
Polycyclic aromatic hydrocarbons (PAHs) are established cancer initiators that can be found in our food and environment. Some dietary plant polyphenols are strong inhibitors to PAH-induced mutagenesis, whereas others may not be as effective. To identify the chemopreventive compounds from a huge volume of dietary components, the development of an efficient screening method is required. In this study, a xenobiotic response element (XRE)-luciferase reporter plasmid was constructed to screen for some potential chemopreventive agents in tea against PAH-induced DNA damage. Tea is one of the most consumed beverages worldwide, and its beneficial effects on health have been documented. Previous studies have claimed that tea polyphenols could be protective against various cancers, and the rich database can be a source for comparison. Among the green and black tea polyphenols, the XRE-luciferase reporter assays suggested that only epigallocatechin gallate (EGCG) was effective in reducing XRE-driven luciferase assay in MCF-7 cells at the concentrations tested. Further study indicated EGCG could reduce CYP1A1 and CYP1B1 mRNA abundances and decrease the DMBA-DNA lesions. The results of DNA covalent binding of all tea polyphenols tested were consistent with the XRE-reporter assays. This study illustrated that the XRE-reporter assay was a viable screening test for dietary chemopreventive agents against PAH-initiated breast mutagenesis. It has the advantages of shorter sample processing time and producing no radioactive waste over directly measuring the CYP1A1/1B1 expressions, DNA lesion, or gel mobility shift assay.
Chan
Ho Yee
HY
Department of Biochemistry, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong.
Wang
Huan
H
Tsang
David S C
DS
Chen
Zhen-Yu
ZY
Leung
Lai K
LK
D000704:Analysis of Variance; D000818:Animals; D001943:Breast Neoplasms / Q000235:genetics / Q000517:prevention & control*; D002470:Cell Survival / Q000187:drug effects; D004249:DNA Damage / Q000187:drug effects; D004353:Drug Evaluation, Preclinical; D005419:Flavonoids / Q000627:therapeutic use*; D017930:Genes, Reporter / Q000502:physiology; D008156:Luciferases; D010636:Phenols / Q000627:therapeutic use*; D011084:Polycyclic Aromatic Hydrocarbons / Q000633:toxicity*; D059808:Polyphenols; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013662:Tea; D014407:Tumor Cells, Cultured / Q000187:drug effects; D015262:Xenobiotics
D005419:Flavonoids; D010636:Phenols; D011084:Polycyclic Aromatic Hydrocarbons; D059808:Polyphenols; D013662:Tea; D015262:Xenobiotics; D008156:Luciferases