KMID : 0379520020180010059
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Çѱ¹µ¶¼ºÇÐȸÁö 2002 Volume.18 No. 1 p.59 ~ p.64
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Screening Assay for Identification of Endocrine Disruptors with Androgen Activities using LNCaP Cells
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Kim Jin-Ho
Chung Hye-Joo Kim Young-Ok Chung Seung-Tae Park Jae-Hyun Cho Dae-Hyun Kim Dong-Sup
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Abstract
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Substantial evidences have been accumulated about the hormone-like effects of exogenous substances such as pesticides and industrial chemicals during past years. The effects of these substances on the endocrine system are believed to be either enhancing or reducing of various endocrine action. It is necessary to identify putative causal agents by the batter system and to assess their ability to disrupt the endocrine system. A variety of in vitro and In vivo approaches have been used to determine the androgenic effects of environmental chemicals. To establish the method for assessment of the putative endocrine disruptors with androgenic activity, we carried out the cell proliferation assay by MTS method after treatment with the various concentration of testosterone in LNCaP cells (human prostatic cancer cell line) and also observed the expression of androgen-related genes by quantitative RT-PCR. In the cell proliferation assay, the results showed that the grouth of LNCaP cells increased within level of at least 10pM testosterone. We measured by quantitative RT-PCR method on the effects of testosterone on mRNA expression of androgen receptor (AR), prostate-specific antigen (PSA), bone morphogenetic protein (BMP) and BMP receptor (BMPR) In LNCaP cells. The results demonstrated that mRNA expression of PSA and BMPR-IB was observed differently within level of at least 0.01 pM testosterone compared with non-treated control. These observations suggest that the detection of PSA and BMPR-IB mRNA by the quantitative RT-PCR in LNCaP cells is very sensitive method to identify the endocrine disruptors to have the androgenic effects.
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KEYWORD
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Endocrine disruptor, Androgen, LNCaP cells, PSA and BMPR-IB, RT-PCR
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