定量分析肝癌形成过程中肝缺氧诱导因子-1α及其异常基因表达。

Quantitative analysis of hepatic hypoxia-inducible factor-1alpha and its abnormal gene expression during the formation of hepatocellular carcinoma.

机构信息

Research Center of Clinical Molecular Biology, Affiliated Hospital of Nantong University, Nantong 226001, China.

出版信息

Hepatobiliary Pancreat Dis Int. 2009 Aug;8(4):407-13.

PMID:19666411

Abstract

BACKGROUND

Hepatic hypoxia-inducible factor-1 (HIF-1) is activated in the progression of hepatocellular carcinoma (HCC). This study aimed to investigate the dynamic alterations of HIF-1alpha and its gene expression so as to explore the relationship between HIF-1alpha expression and hepatocarcinogenesis at the early stage of HCC.

METHODS

A hepatoma model was made with 2-fluorenyl-acetamide (2-FAA) in male Sprague-Dawley rats. Morphological changes of rat hepatocytes were assessed pathologically (HE staining). The dynamic expression of hepatic and circulating HIF-1alpha was quantitatively analyzed by ELISA. The gene fragments of hepatic HIF-1alpha mRNA were amplified by RT-PCR and confirmed by sequencing. The cellular distribution of hepatic HIF-1alpha expression was confirmed by immunohistochemistry.

RESULTS

Histological examination confirmed granule-like degeneration to atypical hyperplasia and HCC development in rat hepatocytes and progressive increases in the levels of hepatic and circulating HIF-1alpha and its gene expression during the course. The levels of HIF-1alpha expression in the liver and blood of rats with hepatoma were significantly higher than those in normal rats and those with degeneration. Immunohistochemical analysis confirmed the positive expression and hepatocyte distribution of HIF-1alpha in the development of rat hepatoma. A positive relationship was found between HIF-1alpha expression in the liver and blood (P<0.01).

CONCLUSIONS

The above observations support the hypothesis that the overexpression of HIF-1alpha and its gene are closely associated with the malignant transformation of hepatocytes and play an important role at the stage of hepatocarcinogenesis.

摘要

背景

肝缺氧诱导因子-1（HIF-1）在肝细胞癌（HCC）的进展中被激活。本研究旨在探讨 HIF-1α的动态变化及其基因表达，以探索 HIF-1α表达与 HCC 早期发生之间的关系。

方法

采用 2-氟乙酰氨基酚（2-FAA）在雄性 Sprague-Dawley 大鼠中建立肝癌模型。通过病理（HE 染色）评估大鼠肝细胞的形态变化。通过 ELISA 定量分析肝内和循环 HIF-1α的动态表达。通过 RT-PCR 扩增肝 HIF-1α mRNA 的基因片段，并通过测序进行确认。通过免疫组织化学法确认肝 HIF-1α表达的细胞分布。

结果

组织学检查证实，大鼠肝细胞出现颗粒状变性、非典型增生和 HCC 发展，肝内和循环 HIF-1α及其基因表达水平在整个过程中逐渐增加。肝癌大鼠肝和血中 HIF-1α的表达水平明显高于正常大鼠和变性大鼠。免疫组织化学分析证实 HIF-1α在大鼠肝癌发生过程中的阳性表达和肝细胞分布。肝内和血中 HIF-1α的表达呈正相关（P<0.01）。