Abstract
Hepatocellular carcinoma is one of the most common tumors in the world. The purpose of the present study was to investigate the inhibitory effects of adenoviral transduction of human melanoma differentiation-associated gene −7 (MDA −7) gene on hepatocellular carcinoma, so as to provide a theoretical basis for gene therapy of the disease. The human MDA −7 gene was cloned into replication-defective adenovirus specific to HepG2 cells using recombinant virus technology. RT-PCR and Western blotting assays were used to determine the expression of human MDA −7 mRNA and MDA −7 protein in HepG2 cells in vitro. Induction of apoptosis by overexpression of the human MDA −7 gene was determined by flow cytometry. In-vivo efficacy of adenoviral delivery of the human MDA −7 gene was assessed in nude mice bearing HepG2 cell lines in vivo by determining inhibition of tumor growth, VEGF and CD34 expression, and microvascular density (MVD). The results showed that AdGFP/MDA −7 induced apoptosis of HepG2 cells in vitro and significantly inhibited tumor growth in vivo (P < 0.05). The intratumoral MVD decreased significantly in the treated tumors (P < 0.05). We conclude the recombination adenovirus AdGFP/MDA −7 can effectively express biologically active human MDA −7, which leads to inhibition of hepatocellular carcinoma growth.
Highlights
Hepatocellular carcinoma (HCC) is one of the most common tumors worldwide
The results demonstrated that the melanoma differentiation-associated gene-7 (MDA-7) cDNA sequence was consistent with that in the GenBank, which indicated that the recombinant adenoviral vector AdvGFP/MDA-7 was constructed successfully
To determine the optimal multiplicity of infection (MOI) for a maximal transgene expression, HepG2 cells were infected with AdvGFP/MDA-7 at various MOIs and examined by fluorescence microscopy
Summary
Hepatocellular carcinoma (HCC) is one of the most common tumors worldwide. Surgical resection is recognized as the most effective method for treatment of HCC. The melanoma differentiation-associated gene-7 (MDA-7) is a novel tumor suppressor gene belonging to the IL-10 cytokine superfamily. It was first identified in 1995 by subtraction hybridization of a cDNA library derived from a human HO-1 melanoma cell line treated with interferon (IFN)-β and protein kinase C activator, mezerin[3]. Overexpression of MDA-7 has been shown to induce tumor cell apoptosis in vitro in a wide variety of cancer cells including melanoma, Pan X et al / Journal of Biomedical Research, 2012, 26(1): 53-58 breast and colorectal cancer[4,5,6]. MDA7-induced apoptosis is tumor selective with minimal cytotoxicity observed in normal human cells. Adenovirus can be used to mediate gene transfer in a wide range of cell types and high level transgene expression can be obtained at high titers[7]
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