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Purpose  To explore expression and function of astrocyte elevated gene-1 ( AEG-1 ) in human retinoblastoma (RB).    Methods  The expression of  AEG-1  in histological sections of human RBs and in RB cell lines was examined using immunohistochemical staining and RT-PCR and Western blotting respectively. We knocked down  AEG-1  gene levels by  AEG-1 -siRNA lentivirus transfection of human RB cell lines SO-RB50 and Y79, and using an MTT assay, we assessed the role of  AEG-1  on RB cell proliferation. The biological significance of lentivirus transfection induced  AEG-1  down-regulation was examined by assessing the apoptosis rate in the transfected RB cells by Annexin V-APC staining and flow cytometry. We additionally measured the expression of Bcl-2, Bax, cleaved-caspase-3 and caspase-3, and the phosphorylation and non-phosphorylation alternation of MAPKs.    Results   AEG-1  expression was detected to be strongly positive in the histological slides of 35 out of 54 (65%) patients with RB.  AEG-1  expression increased significantly ( P <0.05) with tumor stage. In the RB cell lines SO-RB50, Y79 and WERI-RB1 as compared with retinal pigment epithelium cells, expression of  AEG-1  mRNA and AEG-1 protein was significantly higher. In  AEG-1 -siRNA lentivirus transfected cell cultures as compared with negative control lentivirus transfected cell cultures, levels of  AEG-1  mRNA and of AEG-1 protein ( P <0.05) and cell growth rates ( P <0.01) were significantly lower, and apoptosis rate ( P <0.001), Bax/Bcl-2 ratio and cleaved-caspase-3 protein level were significantly increased. The P-ERK/ERK ratio was significantly decreased in the  AEG-1 -siRNA lentivirus transfected cell lines.    Conclusions  Expression of  AEG-1  was associated with RB, in histological slides of patients and in cell culture experiments. Lentivirus transfection induced knockdown of  AEG-1  had a tumor suppressive effect, potentially by tumor cell apoptosis induction through inhibition of ERK.

Lentivirus-Mediated Knockdown of Astrocyte Elevated Gene-1 Inhibits Growth and Induces Apoptosis through MAPK Pathways in Human Retinoblastoma Cells