Comparison of in Vivo and in Vitro Reporter Gene Assays for Short-Term Screening of Estrogenic Activity

AbstractCircular RNAs (circRNAs), continuous loops of single-stranded RNA, regulate gene expression during the development of various cancers. However, the function of circRNAs in hepatocellular carcinoma (HCC) is rarely discussed. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine the mRNA levels of circ_0011385, miR-361-3p, and STC2 in 96 pairs of HCC tissues (tumor tissues and adjacent normal tissues), HCC cell lines, and L02 (human normal liver cell line) cells. The relationships between circ_0011385 expression and clinical features of HCC were evaluated. Functional experiments in vitro or in vivo were used to evaluate the biological function of circ_0011385. Bioinformatics analysis was performed to predict miRNAs and mRNAs sponged by circ_0011385. RNA immunoprecipitation (RIP) and dual-luciferase reporter gene assays were used to elucidate the interactions among circ_0011385, miR-361-3p, and STC2 (stanniocalcin 2). ChIP and dual-luciferase reporter gene assays were used to identify the upstream regulator of circ_0011385. High expression of circ_0011385 was observed in HCC tissues and cell lines and was significantly associated with tumor size, TNM stage, and prognosis. In addition, inhibition of circ_0011385 expression prevented the proliferation of HCC cells in vitro and in vivo. Circ_0011385 sponged miR-361-3p, thereby regulating the mRNA expression of STC2. In addition, the transcription of circ_0011385 was regulated by SP3. Circ_0011385 knockdown suppressed cell proliferation and tumor activity in HCC. Circ_0011385 may therefore serve as a new biomarker in the diagnosis and treatment of HCC.

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Cellular Metabolism in High-Throughput In Vitro Reporter Gene Assays and Implications for the Quantitative In Vitro–In Vivo Extrapolation

Chemical Research in Toxicology ◽

10.1021/acs.chemrestox.0c00037 ◽

2020 ◽

Vol 33 (7) ◽

pp. 1770-1779 ◽

Cited By ~ 2

Author(s):

Fabian C. Fischer ◽

Cedric Abele ◽

Luise Henneberger ◽

Nils Klüver ◽

Maria König ◽

...

Keyword(s):

Reporter Gene ◽

High Throughput ◽

Cellular Metabolism ◽

In Vivo Extrapolation ◽

Reporter Gene Assays

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Estrogenic activity of UV filters determined by an in vitro reporter gene assay and an in vivo transgenic zebrafish assay

Archives of Toxicology ◽

10.1007/s00204-002-0348-4 ◽

2002 ◽

Vol 76 (5-6) ◽

pp. 257-261 ◽

Cited By ~ 104

Author(s):

Richard Schreurs ◽

Peter Lanser ◽

Willem Seinen ◽

Bart van der Burg

Keyword(s):

Reporter Gene ◽

Estrogenic Activity ◽

Reporter Gene Assay ◽

Transgenic Zebrafish ◽

Uv Filters ◽

Gene Assay

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Detection of estrogenic activity in sediment-associated compounds using in vitro reporter gene assays

The Science of The Total Environment ◽

10.1016/s0048-9697(01)01146-9 ◽

2002 ◽

Vol 293 (1-3) ◽

pp. 69-83 ◽

Cited By ~ 122

Author(s):

Juliette Legler ◽

Martine Dennekamp ◽

A.Dick Vethaak ◽

Abraham Brouwer ◽

Jan H Koeman ◽

...

Keyword(s):

Reporter Gene ◽

Estrogenic Activity ◽

Reporter Gene Assays

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Case study in 21 st century ecotoxicology: using in vitro aromatase inhibition data to predict short term in vivo responses in adult female fish

Environmental Toxicology and Chemistry ◽

10.1002/etc.4968 ◽

2020 ◽

Author(s):

Daniel L. Villeneuve ◽

Brett R. Blackwell ◽

Jenna E. Cavallin ◽

Wan‐Yun Cheng ◽

David J. Feifarek ◽

...

Keyword(s):

Adult Female ◽

Female Fish ◽

Aromatase Inhibition ◽

Short Term ◽

Inhibition Data

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A NEW NON-PROTEIN NITROGEN SOURCE FOR RUMINANTS

Canadian Journal of Animal Science ◽

10.4141/cjas69-022 ◽

1969 ◽

Vol 49 (2) ◽

pp. 135-141 ◽

Cited By ~ 2

Author(s):

L. P. Milligan ◽

A. R. Robblee ◽

J. C. Wood ◽

W. C. Kay ◽

S. K. Chakrabartty

Keyword(s):

Nitrogen Source ◽

Dietary Supplement ◽

Nitrogen Retention ◽

Feeding Trial ◽

Short Term ◽

Protein Nitrogen ◽

Rumen Microorganisms ◽

Production Of Ammonia

The preparation of a polymer of urea and furfural containing 23.2% nitrogen is described. This product was converted by rumen microorganisms in vitro to ammonia at a rate approximately one-seventh that of conversion of urea to ammonia. Use of the polymer as a dietary supplement in a feeding trial with lambs improved nitrogen retention over that of unsupplemented controls by 3.45 g of nitrogen retained per day, while an isonitrogenous quantity of supplemental urea improved nitrogen retention by 0.51 g of nitrogen retained per day. The blood urea pattern, throughout the day, of lambs adapted to control, urea-supplemented and urea–furfural polymer-supplemented rations indicated a slow, prolonged production of ammonia from the latter supplement and very rapid, short-term degradation of urea in vivo.

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In vitro and in vivo evaluation of a green fluorescent protein-HSV1-TK dual-function pet reporter gene

Journal of Labelled Compounds and Radiopharmaceuticals ◽

10.1002/jlcr.25804401119 ◽

2001 ◽

Vol 44 (S1) ◽

pp. S339-S341

Author(s):

K. E. Luker ◽

G. D. Luker ◽

C. M. Pica ◽

J. L. Dahlheimer ◽

T. J. Fahrner ◽

...

Keyword(s):

Green Fluorescent Protein ◽

Reporter Gene ◽

Fluorescent Protein ◽

Dual Function ◽

In Vivo Evaluation ◽

Green Fluorescent

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Studies on the functional activity of organotypically cultured mouse ovary

Development ◽

10.1242/dev.15.2.133 ◽

1966 ◽

Vol 15 (2) ◽

pp. 133-141

Author(s):

T. N. Chapekar ◽

G. V. Nayak ◽

Kamal J. Ranadive

Keyword(s):

Functional Activity ◽

Synthetic Medium ◽

Culture Conditions ◽

Short Term ◽

Plasma Clot ◽

Rat Ovary ◽

Old Mice ◽

Term Maintenance

Short-term maintenance of mouse and rat ovary in organotypic culture system is no longer a problem (Martinovitch, 1938; Gaillard, 1953; Trowell, 1959). Gaillard (1953) cultivated ovaries from 7- to 8-day-old and 21-day-old mice for a week on the plasma clot. Trowell (1959) maintained ovaries of 8-day-old mice on a synthetic medium in an O2-CO2 atmosphere for 9 days. He observed no histological differentiation in the tissues of the ovary. What needs confirmation and further investigation is the possibility of maintenance of functional activity of the ovary under culture conditions. A study was therefore undertaken to investigate if an ovary, cultivated in vitro for some time, shows hormonal activity when transplanted in vivo. In the present work cultured ovaries were grafted in the anterior eye-chamber of spayed female mice and the development of secondary sex organs such as mammary glands and uterus was studied.

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Evaluation of cytotoxicity and mutagenicity of the benzodiazepine flunitrazepam in vitro and in vivo

Brazilian Journal of Pharmaceutical Sciences ◽

10.1590/s1984-82502014000200003 ◽

2014 ◽

Vol 50 (2) ◽

pp. 251-256

Author(s):

Igor Vivian de Almeida ◽

Giovana Domingues ◽

Lilian Capelari Soares ◽

Elisângela Düsman ◽

Veronica Elisa Pimenta Vicentini

Keyword(s):

Rattus Norvegicus ◽

Bone Marrow Cells ◽

Micronucleus Test ◽

Term Treatment ◽

Genotoxic Effects ◽

Short Term ◽

Short Term Treatment ◽

Chromosomal Aberration Test

Flunitrazepam (FNZ) is a sedative benzodiazepine prescribed for the short-term treatment of insomnia. However, there are concerns regarding possible carcinogenic or genotoxic effects of this medicine. Thus, the aim of this study was to evaluate the cytotoxic, clastogenic and aneugenic effects of FNZ in hepatoma cells from Rattus norvegicus (HTC) in vitro and in bone marrow cells of Wistar rats in vivo. These effects were examined in vitro following treatment with 0.2, 1.0, 5.0 or 10 μg/mL FNZ using a micronucleus test with a cytokinesis block or in vivo using a chromosomal aberration test following treatment with 7, 15 or 30 μg/mL/kg body weight. The results showed that the benzodiazepine concentrations tested were not cytotoxic, aneugenic or clastogenic. However, considering the adverse effects of using this benzodiazepine, more studies are required.

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