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

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Abstract

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.

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References

  • Alper M, Kockar F (2014) IL-6 upregulates a disintegrin and metalloproteinase with thrombospondin motifs 2 (ADAMTS-2) in human osteosarcoma cells mediated by JNK pathway. Mol Cell Biochem 393(1–2):165–175

    Article  CAS  Google Scholar 

  • Balkwill F (2006) TNF-alpha in promotion and progression of cancer. Cancer Metastasis Rev 25(3):409–416

    Article  CAS  Google Scholar 

  • Desbois-Mouthon C, Cacheux W, Blivet-Van Eggelpoël MJ, Barbu V, Fartoux L, Poupon R, Housset C, Rosmorduc O (2006) Impact of IGF-1R/EGFR cross-talks on hepatoma cell sensitivity to gefitinib. Int J Cancer 119(11):2557–2566

    Article  CAS  Google Scholar 

  • Dodurga Y, Seçme M, Lale Şatıroğlu-Tufan N (2018) A novel oncogene URG4/URGCP and its role in cancer. Gene 668:12–17

    Article  CAS  Google Scholar 

  • Egeblad M, Jäättelä M (2000) Cell death induced by TNF or serum starvation is independent of ErbB receptor signaling in MCF-7 breast carcinoma cells. Int J Cancer 86(5):617–625

    Article  CAS  Google Scholar 

  • Faustman DL, Davis M (2013) TNF receptor 2 and disease: autoimmunity and regenerative medicine. Front Immunol 4:478

    Article  Google Scholar 

  • Haruki K, Shiba H, Fujiwara Y, Furukawa K, Iwase R, Uwagawa T, Misawa T, Ohashi T, Yanaga K (2013) Inhibition of nuclear factor-κB enhances the antitumor effect of tumor necrosis factor-α gene therapy for hepatocellular carcinoma in mice. Surgery 154(3):468–478

    Article  Google Scholar 

  • Jin CY, Park C, Kim GY, Lee SJ, Kim WJ, Choi YH (2009) Genistein enhances TRAIL-induced apoptosis through inhibition of p38 MAPK signaling in human hepatocellular carcinoma Hep3B cells. Chem Biol Interact 180(2):143–150

    Article  CAS  Google Scholar 

  • Kockar FT, Foka P, Hughes TR, Kousteni S, Ramji DP (2001) Analysis of the Xenopus laevis CCAAT-enhancer binding protein alpha gene promoter demonstrates species-specific differences in the mechanisms for both autoactivation and regulation by Sp1. Nucleic Acids Res 29(2):362–372

    Article  CAS  Google Scholar 

  • Kou X, Jin Y, Deng W, Sun K, Han Z, Ye F, Yu G, Fan Q, Gao L, Zhao Q, Zhao X, Li R, Wei L, Wu M (2013) Tumor necrosis factor-α attenuates starvation-induced apoptosis through upregulation of ferritin heavy chain in hepatocellular carcinoma cells. BMC Cancer 13:438

    Article  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  • Li B, Sun C, Sun J, Yang MH, Zuo R, Liu C, Lan WR, Liu MH, Huang B, Zhou Y (2019) Autophagy mediates serum starvation-induced quiescence in nucleus pulposus stem cells by the regulation of P27. Stem Cell Res Ther 10(1):118

    Article  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2 (−Delta Delta C(T)) method. Methods 25(4):402–408

    Article  CAS  Google Scholar 

  • Sabio G, Davis RJ (2014) TNF and MAP kinase signalling pathways. Semin Immunol 26(3):237–245

    Article  CAS  Google Scholar 

  • Satiroglu Tufan NL, Dodurga YD, Gok Cetinkaya A, Feitelson MA (2010) RNA interference-mediated URG4 gene silencing diminishes cyclin D1 mRNA expression in HepG2 cells. Genet Mol Res 9(3):1557–1567

    Article  CAS  Google Scholar 

  • Song J, Xie H, Lian Z, Yang G, Du R, Du Y, Zou X, Jin H, Gao J, Liu J, Fan D (2006) Enhanced cell survival of gastric cancer cells by a novel gene URG4. Neoplasia 8(12):995–1002

    Article  Google Scholar 

  • Tokay E, Kockar F (2016a) Identification of intracellular pathways through which TGF-β1 upregulates URG-4/URGCP gene expression in hepatoma cells. Life Sci 144:121–128

    Article  CAS  Google Scholar 

  • Tokay E, Kockar F (2016b) SP1 is a transcriptional regulator of URG-4/URGCP gene in hepatocytes. Mol Cell Biochem 423(1–2):75–83

    Article  CAS  Google Scholar 

  • Tu CC, Kumar VB, Day CH, Kuo WW, Yeh SP, Chen RJ, Liao CR, Chen HY, Tsai FJ, Wu WJ, Huang CY (2013) Estrogen receptor α (ESR1) over-expression mediated apoptosis in Hep3B cells by binding with SP1 proteins. J Mol Endocrinol 51(1):203–212

    Article  CAS  Google Scholar 

  • Tufan NL, Lian Z, Liu J, Pan J, Arbuthnot P, Kew M, Clayton MM, Zhu M, Feitelson MA (2002) Hepatitis Bx antigen stimulates expression of a novel cellular gene, URG4, that promotes hepatocellular growth and survival. Neoplasia 4(4):355–368

    Article  CAS  Google Scholar 

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Funding

This work was supported by the Balıkesir University Scientific Research Project Grant No. 2017/128.

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Authors and Affiliations

  1. Department of Molecular Biology and Genetic, Faculty of Sciences and Arts, Balıkesir University, Cagis Campus, 10145, Balıkesir, Turkey

    Esra Tokay & Feray Kockar

  2. Department of Medical Biology, Faculty of Medicine, Usak University, Uşak, Turkey

    Rahsan Ilikci Sagkan

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  1. Esra Tokay
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  2. Rahsan Ilikci Sagkan
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Correspondence to Rahsan Ilikci Sagkan or Feray Kockar.

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Tokay, E., Sagkan, R.I. & Kockar, F. TNF-α Induces URG-4/URGCP Gene Expression in Hepatoma Cells through Starvation Dependent Manner. Biochem Genet 59, 300–314 (2021). https://doi.org/10.1007/s10528-020-09972-z

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  • DOI: https://doi.org/10.1007/s10528-020-09972-z

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