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.

URG-4 / URGCP是一个可能与肿瘤发生和细胞周期调控有关的基因。在文献中，没有关于炎性细胞因子介导的URG-4 / URGCP调节的研究。在这项研究中，研究了TNF-α细胞因子对血清饥饿和血清培养的肝癌细胞中URG-4 / URGCP表达的影响。使用MTT和膜联蛋白-V / PI染色以及流式细胞仪分析显示了TNF-α对肝癌细胞的影响。结果，在血清饥饿状态下，TNF-α导致肝癌细胞的细胞毒性，而从血清培养条件下未发现降低。用qRT-PCR分析和Western印迹法在mRNA和蛋白水平上确定TNF-α介导的URG-4 / URGCP表达。在血清饥饿和血清培养的肝癌细胞中，URG-4URGCP mRNA表达均上调。用URG-4 / URGCP启动子构建体进行转染研究，以确定转录活性。TNF-α引起URG / URGCP启动子构建体活性的上调。根据血清条件，URG-4 / URGCP启动子条件的基础活性是不同的。另外，一些通路抑制剂被添加到肝癌细胞中，以阻断特定通路，以发现TNF-α介导的URG-4 / URGCP在mRNA和蛋白质水平上的上调。TNF-α利用JNK和PI3K途径调节血清饥饿的Hep3B细胞中的URG-4 / URGCP基因。在血清培养条件下，渥曼青霉素（PI3K抑制剂），MEK-1（MAPK抑制剂）和SP600125（JNK抑制剂）不抑制TNF-α对URGCP的活化反应。