Candidate dietary phytochemicals modulate expression of phase II enzymes GSTP1 and NQO1 in human lung cells.
Many phytochemicals possess cancer-preventive properties, some putatively through phase II metabolism-mediated mutagen/oxidant quenching. We applied human lung cells in vitro to investigate the effects of several candidate phytopreventive agents, including green tea extracts (GTE), broccoli sprout extracts (BSE), epigallocatechin gallate (EGCG), sulforaphane (SFN), phenethyl isothiocyanate (PEITC), and benzyl isothiocyanate (BITC), on inducing phase II enzymes glutathione S-transferase P1 (GSTP1) and NAD(P)H:quinone oxidoreductase 1 (NQO1) at mRNA and protein levels. Primary normal human bronchial epithelial cells (NHBE), immortalized human bronchial epithelial cells (HBEC), and lung adenocarcinoma cells (A549) were exposed to diet-achievable levels of GTE and BSE (0.5, 1.0, 2.0 mg/L), or individual index components EGCG, SFN, PEITC, BITC (0.5, 1.0, 2.0 micromol/L) for 24 h, 48 h, and 6 d, respectively. mRNA assays employed RNA-specific quantitative RT-PCR and protein assays employed Western blotting. We found that in NHBE cells, while GSTP1 mRNA levels were slightly but significantly increased after exposure to GTE or BSE, NQO1 mRNA increased to 2- to 4-fold that of control when exposed to GTE, BSE, or SFN. Effects on NQO1 mRNA expression in HBEC cells were similar. NQO1 protein expression increased up to 11.8-fold in SFN-treated NHBE cells. Both GSTP1 and NQO1 protein expression in A549 cells were constitutively high but not induced under any condition. Our results suggest that NQO1 is more responsive to the studied chemopreventive agents than GSTP1 in human lung cells and there is discordance between single agent and complex mixture effects. We conclude that modulation of lung cell phase II metabolism by chemopreventive agents requires cell- and agent-specific discovery and testing.
Tan
Xiang-Lin
XL
Division of Pulmonary Medicine, Albert Einstein College of Medicine, Bronx, NY, USA. tan.xianglin@mayo.edu
Shi
Miao
M
Tang
Hui
H
Han
Weiguo
W
Spivack
Simon D
SD
D016588:Anticarcinogenic Agents / Q000494:pharmacology*; D001937:Brassica / Q000737:chemistry; D001980:Bronchi; D028241:Camellia sinensis / Q000737:chemistry; D002392:Catechin / Q000031:analogs & derivatives / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D045744:Cell Line, Tumor; D004847:Epithelial Cells; D015870:Gene Expression / Q000187:drug effects; D051549:Glutathione S-Transferase pi / Q000032:analysis / Q000235:genetics*; D006801:Humans; D017879:Isothiocyanates / Q000494:pharmacology; D008168:Lung / Q000201:enzymology*; D008175:Lung Neoplasms; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000032:analysis / Q000235:genetics*; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013454:Sulfoxides; D013861:Thiocyanates / Q000494:pharmacology
D016588:Anticarcinogenic Agents; D017879:Isothiocyanates; D010936:Plant Extracts; D012333:RNA, Messenger; D013454:Sulfoxides; D013861:Thiocyanates; C058305:phenethyl isothiocyanate; C031403:benzyl isothiocyanate; D002392:Catechin; C045651:epigallocatechin gallate; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human; C496556:GSTP1 protein, human; D051549:Glutathione S-Transferase pi; C016766:sulforaphane