Original Article
Copyright ©2010 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Oct 7, 2010; 16(37): 4677-4684
Published online Oct 7, 2010. doi: 10.3748/wjg.v16.i37.4677
Oncolytic adenovirus SG600-IL24 selectively kills hepatocellular carcinoma cell lines
Xin-Bo Xue, Chao-Wen Xiao, Hui Zhang, Ai-Guo Lu, Wei Gao, Zhu-Qing Zhou, Xin-Lai Guo, Ming-An Zhong, Yao Yang, Cong-Jun Wang
Xin-Bo Xue, Chao-Wen Xiao, Department of Biliary and Pancreatic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China
Hui Zhang, Ai-Guo Lu, Wei Gao, Zhu-Qing Zhou, Xin-Lai Guo, Ming-An Zhong, Yao Yang, Cong-Jun Wang, Department of General Surgery, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200120, China
Author contributions: Xiao CW and Zhang H contributed equally to this work; Zhang H and Wang CJ designed the research; Guo XL, Zhong MA and Yang Y provided the vital reagents; Lu AG, Gao W and Zhou ZQ collected and analyzed the data; Xiao CW wrote the manuscript and performed the majority of experiments; Xue XB and Wang CJ reviewed the manuscript.
Supported by National Natural Science Foundation of China, No. 30872510; Natural Science Foundation of Hubei Province, No. 2008CDB127
Correspondence to: Cong-Jun Wang, MD, PhD, Department of General Surgery, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200120, China. wcj902@163.com
Telephone: +86-21-38804518 Fax: +86-21-58765589
Received: June 7, 2010
Revised: July 5, 2010
Accepted: July 12, 2010
Published online: October 7, 2010
Abstract

AIM: To investigate the effect of oncolytic adenovirus SG600-IL24 and replication-incompetent adenovirus Ad.IL-24 on hepatocellular carcinoma (HCC) cell lines and normal liver cell line.

METHODS: HCC cell lines (HepG2, Hep3B and MHCC97L) and normal liver cell line (L02) with a different p53 status were infected with SG600-IL24 and Ad.IL-24, respectively. Melanoma differentiation-associated (MDA)-7/interleukin (IL)-24 mRNA and protein expressions in infected cells were detected by reverse transcription-polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and Western blotting, respectively. Apoptosis of HCC cells and normal liver cells was detected by cytometric assay with Hoechst33258 staining. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to investigate proliferation of HCC cells and normal liver cells, and cell cycle was assayed by flow cytometry.

RESULTS: RT-PCR, ELISA and Western blotting showed that the exogenous MDA-7/IL-24 gene was highly expressed in cells infected with SG600-IL24. MTT indicated that SG600-IL24 could suppress the growth of HepG2, Hep3B, MHCC97L, with an inhibition rate of 75% ± 2.5%, 85% ± 2.0%, 72% ± 1.8%, respectively (P < 0.01), promote the apoptosis of HepG2, Hep3B, MHCC97L, with an apoptosis rate of 56.59% ± 4.0%, 78.36% ± 3.5%, 43.39% ± 2.5%, respectively (P < 0.01), and block the HCC cell lines in the G2/M phase with a blocking rate of 35.4% ± 4.2%, 47.3% ± 6.2%, 42% ± 5.0%, respectively (P < 0.01) but not the normal liver cell line in a p53-independent manner.

CONCLUSION: SG600-IL24 can selectively suppress the proliferation and apoptosis of HCC cell lines in vitro but not normal liver cell line L02 in a p53-independent manner. Compared with Ad.IL-24, SG600-IL24 can significantly enhance the antitumor activity in HCC cell lines.

Keywords: Oncolytic adenovirus; Hepatocellular carcinoma; Cancer gene therapy; p53-independent; Melanoma differentiation-associated-7/interleukin-24