Abstract

Hepatocellular Carcinoma (HCC) is the fourth leading cause of cancer-related death in the World. Epidermal Growth Factor Receptor (EGFR) pathway plays an important role in HCC tumorigenesis. In the tumor microenvironment of HCC, the expression of EGF is aberrant. Here we describe the EGF-dependent regulation of URGCP gene in Human Hepatoma cancer cells (Hep3B). The effect of EGF cytokine on Hep3B proliferation was shown using MTT method. EGF-mediated URGCP expression was determined at mRNA and protein level with qRT-PCR analyses and Western blotting method, respectively. Different lengths of URGCP promoter constructs were transient transfected in to Hep3B cells and the basal promoter activities were determined in the presence of EGF. In addition, some pathway analyses were performed to find out the mechanism of EGF mediated up-regulation of the URGCP gene. In the presence of EGF, URGCP expression increases in concentration and time dependent manner. EGF mediated URGCP up-regulation might depend on a cis-acting element located between –344 and –482 positions in its promoter.

中文翻译：

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表皮生长因子介导人肝癌细胞中 URGCP 癌基因的上调

摘要

肝细胞癌 (HCC) 是全球癌症相关死亡的第四大原因。表皮生长因子受体 (EGFR) 通路在 HCC 肿瘤发生中起重要作用。在HCC的肿瘤微环境中，EGF的表达异常。在这里，我们描述了URGCP的 EGF 依赖性调节人肝癌细胞 (Hep3B) 中的基因。使用MTT方法显示EGF细胞因子对Hep3B增殖的影响。分别用 qRT-PCR 分析和蛋白质印迹法在 mRNA 和蛋白质水平测定 EGF 介导的 URGCP 表达。将不同长度的 URGCP 启动子构建体瞬时转染到 Hep3B 细胞中，并在 EGF 存在下测定基础启动子活性。此外，还进行了一些途径分析以找出 EGF 介导的 URGCP 基因上调的机制。在 EGF 存在下，URGCP表达以浓度和时间依赖性方式增加。EGF 介导的URGCP上调可能取决于位于其启动子中 –344 和 –482 位置之间的顺式作用元件。