title: Quantitative analysis of hepatic hypoxia-inducible factor-1alpha and its abnormal gene expression during the formation of hepatocellular carcinoma.
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.
authors: Research Center of Clinical Molecular Biology, Affiliated Hospital of Nantong University, Nantong 226001, China. yaodf@ahnmc.com
authors: Deng-Fu
authors: DF
authors: Yao
authors: Hua
authors: H
authors: Jiang
authors: Min
authors: M
authors: Yao
authors: Yue-Ming
authors: YM
authors: Li
authors: Wen-Jing
authors: WJ
authors: Gu
authors: Yu-Cheng
authors: YC
authors: Shen
authors: Li-Wei
authors: LW
authors: Qiu
authors: Wei
authors: W
authors: Wu
authors: Xin-Hua
authors: XH
authors: Wu
authors: Wen-Li
authors: WL
authors: Sai
mesh_terms: D015073:2-Acetylaminofluorene; D000818:Animals; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D002471:Cell Transformation, Neoplastic / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D015972:Gene Expression Regulation, Neoplastic; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008113:Liver Neoplasms / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008969:Molecular Sequence Data; D011230:Precancerous Conditions / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
chemical_list: C497444:Hif1a protein, rat; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012333:RNA, Messenger; D015073:2-Acetylaminofluorene