Abstract 72: Analyzing the role of Astrocyte Elevated Gene-1 (AEG-1) in hepatocarcinogenesis using a knockout mouse model
Authors: Robertson Chadia L. , Srivastava Jyoti , Siddiq Ayesha , Gredler Rachel , Rajasekaran Devaraja , Akiel Maaged , Shen Xue-Ning , Arkun Knarik , Ghosh Shobha , Subler Mark A. , Windle Jolene , Fisher Paul B. , Sarkar Devanand 
Published: May 2015

The oncogene Astrocyte Elevated Gene-1 (AEG-1) is overexpressed in a wide variety of human cancers. In vitro and in vivo nude mouse xenograft studies have established the tumor-promoting role of AEG-1. Activation of multiple signaling pathways, such as NF-κB, PI3K/Akt, Wnt/β-catenin and MEK/ERK, has been attributed to mediate oncogenesis by AEG-1. To better understand the role AEG-1 plays in the onset and progression of cancer and to identify key mechanism(s) of AEG-1 action, we generated a novel AEG-1 knockout mouse (AEG-1-/-). We studied hepatocarcinogenesis in this model because AEG-1 is overexpressed in more than 90% of human hepatocellular carcinoma (HCC) patients and is believed to play a key regulatory role in progression of the disease. AEG-1-/- mice were viable with normal development. We first compared the aging associated phenotype of WT and AEG-1-/- mice. At 16 months of age spontaneous tumor development in lungs and liver was observed in some cohorts of untreated WT mice (n=20). Such tumorigenesis was not detected in any of the AEG-1-/- mice. Additionally, age-associated inflammatory infiltrates were observed in the internal organs of WT but not AEG-1-/- mice. To initiate HCC, WT and AEG-1-/- mice were treated with diethylnitrosamine (DEN) at 2 weeks of age. At 32 weeks WT mice showed a marked hepatocarcinogenic response while AEG-1-/- mice exhibited profound resistance to tumor formation. The hepatocarcinogenic response was further evaluated by DEN initiation followed by daily phenobarbital treatment. WT mice exhibited an intensified hepatocarcinogenic response evidenced by large necrotic liver tumors at 28 weeks of age with a 52% rate of lung metastasis. AEG-1-/- mice remained remarkably resistant even to this combinatorial treatment with no distant metastasis. No difference in the activation of Akt, ERK and β-catenin signaling was observed between WT and AEG-1-/- mice indicating that these pathways may not be regulated by AEG-1 under physiological conditions. However, marked inhibition of NF-κB activation was observed in AEG-1-/- mice. Lipopolysaccharide (LPS)-induced NF-κB luciferase reporter activity, nuclear translocation of p65 subunit of NF-κB and induction of IL-1β and IL-6 were markedly attenuated in primary hepatocytes and macrophages isolated from AEG-1-/- versus WT mice. Chronic inflammation and NF-κB activation play a causal role in hepatocarcinogenesis. Inhibition of NF-κB activation in hepatocytes and macrophages profoundly abrogates HCC development in mouse models. Our findings in AEG-1-/- mice confirm a fundamental role of AEG-1 in NF-κB activation. Here we demonstrate that lack of AEG-1 protects from generalized inflammation thereby suppressing spontaneous as well as experimental tumorigenesis. AEG-1-/-mice might be a useful model to study inflammation and cancer in diverse organ systems.

Citation Format: Chadia L. Robertson, Jyoti Srivastava, Ayesha Siddiq, Rachel Gredler, Devaraja Rajasekaran, Maaged Akiel, Xue-Ning Shen, Knarik Arkun, Shobha Ghosh, Mark A. Subler, Jolene Windle, Paul B. Fisher, Devanand Sarkar. Analyzing the role of Astrocyte Elevated Gene-1 (AEG-1) in hepatocarcinogenesis using a knockout mouse model. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 72. doi:10.1158/1538-7445.AM2014-72