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KMID : 0613820190290101159
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Journal of Life Science
2019 Volume.29 No. 10 p.1159 ~ p.1163
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Establishment of an In Vitro TCD (Testosterone Compound Detection) System
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Lee Dong-Geun
Jo Jung-Kwon
Lee Sang-Hyeon
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Abstract
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Although there is a growing interest in male menopause, a phenomenon associated with male hormone depletion, current kits using antibodies to quantify male hormones are expensive. In this study, we constructed an in vitro system for verifying the activity or concentration of male steroid hormones using a transcriptional activity test. A reporter plasmid, pGL2-Neo-ARE-AdE1BTATA, which reacts to testosterone, was constructed. In this plasmid, the ARE-AdE1bTATA sequences can be bounded by the testosterone - androgen receptor complex to express luciferase as a reporter. Then, a stable transfection was performed on the human prostate cancer cell line, LNcap-LN3. The constructed LNcap- LN3/pGL2-Neo-ARE-AdE1BTATA testosterone compound detection (TCD) system showed quantitatively proportional luciferase activities to concentrations of 10-13 to 10-8 M of standard testosterone. The established in vitro TCD system will contribute to the development of materials for health/functional foods and drugs as it will be possible to search en masse for testosterone-like or testosterone-inhibiting substances derived from natural materials.
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KEYWORD
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Androgen, luciferase, reporter, testosterone
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