Establishment of a fluorescence resonance energy transfer-based bioassay for detecting dioxin-like compounds

Lin, Chi‐Iou; Hsieh, Chun‐Hung; Lee, Stanley Shiao-Ying; Lee, Wei-Shan; Chang-Chien, Gou-Ping; Pan, Chien‐Yuan; Lee, Hsinyu

doi:10.1007/s11373-008-9267-6

Cited by 7 publications

(3 citation statements)

References 39 publications

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“…The pGFP‐C1‐AhR vector was obtained from Dr. Hsin‐yu Lee (Life Science, National Taiwan University, Taipei, Taiwan; ref. 21). At 24 h after transfection, cells were cultured on coverslips and incubated overnight, followed by incubation in serum‐free medium for 24 h. The cells were then stimulated with phthalates and subjected to continuous green fluorescent protein (GFP) analysis by TIRF microscopy.…”

Section: Total Internal Reflection Fluorescence (Tirf) Microscopymentioning

confidence: 99%

Phthalates induce proliferation and invasiveness of estrogen receptor‐negative breast cancer through the AhR/HDAC6/c‐Myc signaling pathway

Hsieh¹,

Tsai²,

Hsu³

et al. 2011

FASEB j.

156

102

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The environmentally present group of chemical phthalates, or phthalate esters, has been recognized as a rising threat to public health, including cancer. While most studies have addressed the estrogenic effects of phthalates in malignancies of the breast and the prostate, little is known about their role in the etiology of hormone-independent cancer. Here we show that treatments with the phthalates n-butyl benzyl phthalate (BBP) and dibutyl phthalate (DBP) at 1 μM induced proliferation (BBP, 3.2-fold; DBP, 3.2-fold), migration (BBP, 2.6-fold; DBP, 2.6-fold), invasion (BBP, 2.7-fold; DBP, 3.1-fold), and tumor formation (EC(50): BBP, 0.12 μM; DBP, 0.22 μM) in estrogen receptor (ER)-negative breast cancer cells (MDA-MB-231). We further demonstrate that phthalates stimulated the cell surface aryl hydrocarbon receptor (AhR) and triggered the downstream cyclic AMP (cAMP)-PKA-CREB1 signaling cascade. The pathway led to increased expression of HDAC6, which facilitated nuclear assembly of the β-catenin-LEF1/TCF4 transcriptional complex and transactivation of the c-Myc oncogene. This nongenomic pathway emanated from the phthalate-induced AhR promoted tumorigenesis of ER-negative breast cancer. Collectively, our findings revealed a novel oncogenic mechanism of phthalates in breast cancer independent from their estrogenic activities.

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Section: Total Internal Reflection Fluorescence (Tirf) Microscopymentioning

confidence: 99%

Phthalates induce proliferation and invasiveness of estrogen receptor‐negative breast cancer through the AhR/HDAC6/c‐Myc signaling pathway

Hsieh¹,

Tsai²,

Hsu³

et al. 2011

FASEB j.

156

102

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“…Human AhR (hAhR) and ARNT were respectively fused to the cyan fluorescent protein (as an energy donor) and the yellow fluorescent protein (YFP, as an energy acceptor), cotransfected into the H4IIEC3 rat hepatoma cell line, and the FRET signal was used to monitor the formation of AhR/ARNT complexes. Hence, a nuclear FRET signal was induced by POPs in a dose‐dependent manner, allowing the detection of 1 p m of TCDD after a 1 hour incubation (Lin et al, ). However, to our knowledge, the highest reported sensitivity among all known bioassays was achieved using BRET and a hAhR fused to the Renilla luciferase (as an energy donor) and a partner consisting of ARNT, heat shock protein 90 or ARNT fused to the energy acceptor YFP (Wang, Liao et al, ).…”

Section: Exogenous Aryl Hydrocarbon Receptor‐based Bioassays: Expandimentioning

confidence: 99%

Aryl hydrocarbon receptor‐based bioassays for dioxin detection: Thinking outside the box

Otarola

Castillo

Marcellini

2017

J of Applied Toxicology

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Despite intensive media coverage and international regulations, man-made persistent organic pollutants such as dioxins represent a serious environmental and health threat. Their detection by sophisticated chromatography technologies is highly complex, impeding the constant monitoring of food or environmental samples. This limitation has fostered the development of generations of bioassays exploiting the molecular function of the aryl hydrocarbon receptor (AhR), which binds toxic compounds and directly activates the transcription of target genes. Here, we review the rich panel of available AhR-dependent bioassays and propose a novel classification based on the source of AhR, which can either be endogenously produced by cell types or tissues naturally responsive to dioxins, or exogenously introduced into a wide range of cellular contexts. In both cases, in vitro and in vivo strategies have been engineered to monitor the formation of molecular complexes, and the activation of direct downstream targets or reporter genes. We evaluate and compare bioassays based on exogenous and endogenous AhR proteins and discuss their specific challenges, strengths and opportunities for futures applications. Undoubtedly, the dynamic field of AhR-dependent bioassays will keep providing new and original strategies to help protect human health and ecosystems from persistent organic pollutants.

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“…The fluorescence resonance energy transfer (FRET) technique can precisely evaluate interaction between two molecules in cells, and the use of cyan-fluorescent protein (CFP) as an energy donor and yellow-fluorescent protein (YFP) as an energy acceptor has been reported as the most suitable combination of FRET signal detection [38]. Lin et al [39] therefore established a FRET-based dioxin-detection assay. Aryl hydrocarbon receptor (AHR) and AHR nuclear translocate (ARNT) fused-cyan fluorescent protein 9CFP) and-yellow fluorescent protein (YFP) constructs were transiently cotransfected into rat hepatoma cell line H4IIEC3 cells.…”

Section: Dioxin Detection By Fret Techniquementioning

confidence: 99%

Biomedical vignette

2008

J Biomed Sci

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Establishment of a fluorescence resonance energy transfer-based bioassay for detecting dioxin-like compounds

Cited by 7 publications

References 39 publications

Phthalates induce proliferation and invasiveness of estrogen receptor‐negative breast cancer through the AhR/HDAC6/c‐Myc signaling pathway

Phthalates induce proliferation and invasiveness of estrogen receptor‐negative breast cancer through the AhR/HDAC6/c‐Myc signaling pathway

Aryl hydrocarbon receptor‐based bioassays for dioxin detection: Thinking outside the box

Biomedical vignette

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