Department of Gastroenterology, Second Xiangya Hospital of Central South University, People's Road 139, Changsha, 410011, China.
Diagn Pathol. 2010 Sep 16;5:58. doi: 10.1186/1746-1596-5-58.
Hepatoma-derived growth factor (HDGF) is involved in the hepatocarcinogenesis. In this study, we investigated the HDGF expression in hepatocellular carcinoma (HCC) and its correlation with clinicopathologic features, including the survival of patients with HCC. Furthermore, we examined the biological processes regulated by HDGF during the development of using HepG2 cell line as a model system.
We used immunohistochemistry to compare HDGF protein expression in HCC and normal liver tissues and further analyze the HDGF protein expression in clinicopathologically characterized 137 HCC cases. We stably knocked down the endogenous expression level of HDGF in HepG2 cells with specific shRNA-expressing lentiviral vector. Following the successful establishment of stable cells, we examined in vitro cell growth by MTT assay, anchorage-independent growth by soft-agar colony formation assay and cell migration/invasion by transwell and boyden chamber assay. And in addition, we also investigated the in vivo tumor growth by xenograft transplantation of HepG2 cells into nude mice.
Protein expression level of HDGF was markedly higher in HCC tissues than that in the normal liver tissues(P = 0.011). In addition, high expression of HDGF protein was positively correlated with T classification(p < 0.001), N classification (p < 0.001), and clinical stage (p < 0.001) of HCC patients. Patients with higher HDGF expression showed a significantly shorter overall survival time than did patients with low HDGF expression. Multivariate analysis suggested that HDGF expression might be an independent prognostic indicator(p < 0.001) for the survival of patients with HCC. HDGF-specific shRNA (shHDGF) successfully knocked down its endogenous expression in HepG2 cells. Compared to the parental and control shRNA-transfected (shCtrl) HepG2 cells, the shHDGF cells exhibited significantly reduced in vitro cell growth, anchorage-independent growth, cell migration and invasion (p < 0.05). In vivo, the xenograft transplants from shHDGF cells gave rise to much smaller tumors as compared to those from shCtrl cells.
High HDGF expression is associated with poor overall survival in patients with HCC. Down-regulation of HDGF inhibits the growth, anchorage-independent growth, migration and invasion of HepG2 cells.
肝癌衍生生长因子（HDGF）参与肝癌的发生。本研究旨在探讨 HDGF 在肝癌中的表达及其与临床病理特征的关系，包括肝癌患者的生存情况。此外，我们还使用 HepG2 细胞系作为模型系统，研究了 HDGF 在肝癌发展过程中调节的生物学过程。
我们使用免疫组织化学方法比较了肝癌组织和正常肝组织中 HDGF 蛋白的表达，并进一步分析了 137 例具有临床病理特征的肝癌病例中 HDGF 蛋白的表达。我们使用特异性 shRNA 表达的慢病毒载体稳定敲低 HepG2 细胞内源性 HDGF 的表达水平。在成功建立稳定细胞后，我们通过 MTT 测定法检测体外细胞生长、软琼脂集落形成测定法检测无锚定生长、Transwell 和 Boyden 室测定法检测细胞迁移/侵袭。此外，我们还通过将 HepG2 细胞异种移植到裸鼠中来研究体内肿瘤生长。
HDGF 蛋白的表达水平在肝癌组织中明显高于正常肝组织（P = 0.011）。此外，HDGF 蛋白的高表达与 HCC 患者的 T 分类（p < 0.001）、N 分类（p < 0.001）和临床分期（p < 0.001）呈正相关。高 HDGF 表达的患者总生存时间明显短于低 HDGF 表达的患者。多变量分析表明，HDGF 表达可能是 HCC 患者生存的独立预后指标（p < 0.001）。HDGF 特异性 shRNA（shHDGF）成功敲低了 HepG2 细胞内源性表达。与亲本和对照 shRNA 转染（shCtrl）的 HepG2 细胞相比，shHDGF 细胞的体外细胞生长、无锚定生长、细胞迁移和侵袭能力显著降低（p < 0.05）。体内实验中，与 shCtrl 细胞相比，shHDGF 细胞的异种移植瘤体积明显较小。
HDGF 高表达与 HCC 患者总体生存不良相关。下调 HDGF 抑制 HepG2 细胞的生长、无锚定生长、迁移和侵袭。
