Alpha-fetoprotein acts as a novel signal molecule and mediates transcription of Fn14 in human hepatocellular carcinoma

• Published in: Journal of hepatology 2012-08, Vol.57 (2), p.322-329
• Main Authors: Wang, Shanshan, Jiang, Wei, Chen, Xiangmei, Zhang, Chao, Li, Hui, Hou, Wenting, Liu, Zhongmin, McNutt, Michael A, Lu, Fengmin, Li, Gang
• Format: Article
• Language: English
• Subjects: Active Transport, Cell Nucleus; All trans retinoic acid; alpha-Fetoproteins - physiology; Base Sequence; Biological and medical sciences; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - metabolism; Fn14; Gastroenterology and Hepatology; Gastroenterology. Liver. Pancreas. Abdomen; Hep G2 Cells; Hepatoma; Humans; Liver Neoplasms - genetics; Liver Neoplasms - metabolism; Liver. Biliary tract. Portal circulation. Exocrine pancreas; Medical sciences; Molecular Sequence Data; Nuclear receptor; Receptors, Retinoic Acid - physiology; Receptors, Tumor Necrosis Factor - genetics; Signal Transduction; Transcription regulation; Transcription, Genetic; Tumors; TWEAK Receptor
• ISSN: 0168-8278; 1600-0641
• DOI: 10.1016/j.jhep.2012.03.029

Background & Aims The function of cytoplasmic AFP as a regulatory factor in the growth of tumor cells has been well defined. However, its precise mechanism of action and its clinical significance remain to be worked out. Methods Specimens from HCC patients were analyzed by using immunohistochemistry, co-immunoprecipitation (CoIP), and chromatin immunoprecipitation (ChIP) assays to evaluate the role of AFP in RAR signaling-mediated carcinogenesis. Quantitative real-time reverse transcription PCR, Western blotting, confocal microscopy, CoIP, GST pull-down, siRNA, gene transfection, and ChIP assays were also used for analysis of cell lines. Results RAR is able to interact with cytoplasmic AFP and binds to the element of the regulatory region of the Fn14 gene in the neoplastic tissue of HCC patients. An assay of hepatocyte cell lines of differing AFP expression showed that cytoplasmic AFP is able to block ATRA-induced nuclear translocation of RAR and expression of the Fn14 gene. Knockdown of AFP in siRNA-transfected HepG2 and Bel7402 cells led to greater binding of RAR to its response element. The expression of the Fn14 gene was therefore up regulated as reflected by increases in mRNA and protein levels. Conversely, transfection of HLE and L02 cells (AFP negative) with the afp gene resulted in apparent reduction of RAR binding to DNA and Fn14 protein. Conclusions Demonstration of the involvement of cytoplasmic AFP in RAR-mediated expression of the Fn14 gene strongly indicates AFP plays a signal molecule-like role in the regulation of hepatocellular carcinoma growth.