Tan Wenting,Luo Xiao,Yan Zehui,Dan Yunjie,Mao Qing,Deng Guohong.ERC-PCR based detection for environmental endocrine-disrupting compounds[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(01):1-4.
Tan Wenting,Luo Xiao,Yan Zehui,Dan Yunjie,Mao Qing,Deng Guohong.ERC-PCR based detection for environmental endocrine-disrupting compounds[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(01):1-4.DOI:
Objective To develop a rapid and high-throughput screening system to detect trace environmental endocrine disrupting compounds (EDCs) in vitro based on the estrogen-dependent binding of estrogen receptors α (ERα) with estrogen response element (ERE). Methods Two DNA sequences
COX7RP and VitA2 gene promoter which contained ERE and were of high affinity to the ERα
were used as combination probes to measure the es
trogenic concentration in the estrogen-dependent ERα-ERE binding process. The combination probe was detected by real-time fluorescent quantitative PCR. Five compounds
17β-estradiol (E
2
)
estrone (E
1
)
diethylstilbestrol (DES) and dimethylphthalate (DMP) at the final concentration of 10
-14
to 10
-6
mol/L were detected by the present method. Results Compared with the VitA2-ERE
human-derived COX7RP-ERE had a better combination with the probe during estrogen-dependent ERα-ERE binding process in vitro. Our present detection system provided sensitive estrogen-dependent dose-response curves for E
1
E
2
DES and DMP
with median effective concentration (EC
50
) at 0.89 nmol/L
19.9 pmol/L
0.79 nmol/L
and 6.31 pmol/L
respectively. Conclusion Our detection system based on ERC-PCR strategy is a sensitive