Methods of analysis for chemicals that disrupt cellular signaling pathways: risk assessment for potential endocrine disruptors.
Here we present a basic concept and several examples of methods of analysis for chemicals that disrupt cellular signaling pathways, in view of risk assessment for potential endocrine disrupting chemicals (EDCs). The key cellular signaling pathways include 1) ER/coactivator interaction, 2) AR translocation into the nucleus, 3) ER/NO/sGC/cGMP, 4) ER/Akt, 5) ER/Src, 6)ER/Src/Grb2, and 7) ER/Ca2+/CaM/CaMK pathways. These were visualized in relevant live cells using newly developed fluorescent and bioluminescent probes. Changes in cellular signals were thereby observed in nongenomic pathways of steroid hormones upon treatment of the target cells with steroid hormones and related chemicals. This method of analysis appears to be a rational approach to high-throughput prescreening (HTPS) of biohazardous chemicals, EDCs, in particular. Also described was the screening of gene expression by serial analysis of gene expression and gene chips upon applying EDCs to breast cancer cells, mouse livers, and human neuroblastoma NB-1 cells.
Umezawa
Yoshio
Y
Department of Chemistry, School of Science, The University of Tokyo, Hongo, Tokyo 113-0033, Japan. umezawa@chem.s.u-tokyo.ac.jp
Ozawa
Takeaki
T
Sato
Moritoshi
M
Inadera
Hidekuni
H
Kaneko
Shuichi
S
Kunimoto
Manabu
M
Hashimoto
Shin-ichi
S
D000818:Animals; D001943:Breast Neoplasms / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D004703:Endocrine System / Q000187:drug effects* / Q000502:physiology; D005456:Fluorescent Dyes; D020869:Gene Expression Profiling; D006801:Humans; D008099:Liver / Q000473:pathology; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D009447:Neuroblastoma / Q000473:pathology; D018570:Risk Assessment; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology; D013256:Steroids / Q000633:toxicity*; D014407:Tumor Cells, Cultured
D005456:Fluorescent Dyes; D013256:Steroids