TNF-α Induces URG-4/URGCP Gene Expression in Hepatoma Cells through Starvation Dependent Manner

URG-4/URGCP is a gene that may be associated with the onset of tumorigenesis and cell cycle regulation. In the literature, there is no study about inflammatory cytokine-mediated URG-4/URGCP regulation. In this study, the effect of TNF-α cytokine was investigated on URG-4/URGCP expression in serum-st...

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Bibliographic Details
Published in:Biochemical genetics 2021-02, Vol.59 (1), p.300-314
Main Authors: Tokay, Esra, Sagkan, Rahsan Ilikci, Kockar, Feray
Format: Article
Language:English
Subjects:
1-Phosphatidylinositol 3-kinase
Biochemistry
Biomedical and Life Sciences
Biomedicine
Cell cycle
Cytokines
Cytotoxicity
Gene expression
Hepatoma
Human Genetics
Inflammation
Inhibitors
Liver cancer
MAP kinase
Medical Microbiology
Original Article
Proteins
Toxicity
Transcription
Transfection
Tumor necrosis factor-α
Tumorigenesis
Western blotting
Wortmannin
Zoology
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Summary:URG-4/URGCP is a gene that may be associated with the onset of tumorigenesis and cell cycle regulation. In the literature, there is no study about inflammatory cytokine-mediated URG-4/URGCP regulation. In this study, the effect of TNF-α cytokine was investigated on URG-4/URGCP expression in serum-starved and serum-cultured hepatoma cells. The effect of TNF-α on hepatoma cells was shown using MTT and Annexin-V/PI staining with flow cytometer analyses. As a result, TNF-α leads to the cytotoxicity of hepatoma cells in serum-starved condition whereas no decrease was detected from serum-cultured condition. TNF-α-mediated URG-4/URGCP expression was determined at mRNA and protein level with qRT-PCR analyses and Western blotting method. URG-4URGCP mRNA expression was upregulated in both serum-starved and serum-cultured hepatoma cells. The transfection studies were carried out with URG-4/URGCP promoter constructs for determining the transcriptional activity. TNF-α caused to the upregulation of the activities of URG/URGCP promoter constructs. The basal activities of the URG-4/URGCP promoter conditions are differential according to serum conditions. In addition, some pathway inhibitors were added into hepatoma cells for blocking specific pathways to find out TNF-α-mediated URG-4/URGCP upregulation at mRNA and protein level. TNF-α used JNK and PI3K pathways for regulating URG-4/URGCP gene at serum-starved Hep3B cells. In serum-cultured condition, wortmannin (PI3K inhibitor), MEK-1 (MAPK inhibitor), and SP600125 (JNK inhibitor) did not inhibit the activation response of TNF-α on URGCP.
ISSN:0006-2928
1573-4927
DOI:10.1007/s10528-020-09972-z