Melanoma differentiation-associated gene-7, MDA-7/IL-24, selectively induces growth suppression, apoptosis in human hepatocellular carcinoma cell line HepG2 by replication-incompetent adenovirus vector

• Published in: World journal of gastroenterology : WJG Vol. 12; no. 11; pp. 1774 - 1779
• Main Authors: Wang, Cong-Jun, Xue, Xin-Bo, Yi, Ji-Lin, Chen, Kun, Zheng, Jian-Wei, Wang, Jian, Zeng, Jian-Ping, Xu, Rong-Hua
• Format: Journal Article
• Language: English
• Published: United States Department of Biliary and Pancreatic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China 21.03.2006; Baishideng Publishing Group Co., Limited
• Subjects: Adenoviridae - genetics; Apoptosis - genetics; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - pathology; Carcinoma, Hepatocellular - therapy; Cell Cycle - genetics; Cell Line, Tumor; Cell Proliferation; Cell Survival - genetics; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Gene Expression Regulation, Neoplastic; Gene Expression Regulation, Viral; Genetic Therapy; Genetic Vectors; Hepatocytes; Humans; Interleukins - genetics; Liver Neoplasms - genetics; Liver Neoplasms - pathology; Liver Neoplasms - therapy; Microscopy, Fluorescence; Rapid Communication; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Neoplasm - genetics; RNA, Viral - genetics; Transfection; Virus Replication; Growth suppression; MDA-7/IL-24; Hepatocellular carcinoma (HCC); Cancer gene therapy; Apoptosis
• ISSN: 1007-9327; 2219-2840
• DOI: 10.3748/wjg.v12.i11.1774

To investigate the effect of replication-incompetent adenovirus vector expressing MDA-7/IL-24 on tumor growth and apoptosis in human hepatocellular carcinoma (HCC) cell line HepG2 and normal liver cell line L02. We constructed the recombinant replication-incompetent Ad.mda-7 virus vector and infected it into the human HCC cell line HepG2 and normal liver cell line L02. RT-PCR was performed to detect the mRNA expressing in cells. by ELISA was used to detect MDA-7/IL-24 protein expression in the culture supernatant. The effect of apoptosis induced by Ad.mda-7 was confirmed by Hoechst staining and flow cytometry assay with Annexin-V and PI staining. MTT assay was used to determine growth inhibition of HepG2 cells, and cell-cycle and hypodiploidy analyses were performed by flow cytometry. Recombinant replication-defective virus expressing MDA-7/IL-24 was constructed successfully. RT-PCR showed that the Ad.mda-7 could mediate the expression of the exogenous gene MDA-7/IL-24 into HepG2 and L02. The concentration of MDA-7/IL-24 protein in supernatant was 130 pg/mL and 110 pg/mL in Ad.mda-7-infected L02 and HepG2 cells, respectively. Ad.mda-7 infection obviously induced apoptosis (from 2.60%+/-0.72% to 33.6%+/-13.2%, P=0.00012) and growth suppression in HepG2 (inhibition ratio IR=68%) and an increase in the percentage of specific cancer cell types at the G2/M phase of the cell cycle (from 6.44% to 32.29%, P<0.01), but not in L02 cells. These results confirm selectively induction of apoptosis and growth suppression by the mda-7/IL-24 gene with replication-incompetent adenovirus vector in human hepatocellular carcinoma cell line HepG2.