Oncolytic adenovirus-mediated MDA-7/IL-24 overexpression enhances antitumor activity in hepatocellul

来源 :Hepatobiliary & Pancreatic Diseases International | 被引量 : 0次 | 上传用户:lele3383
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BACKGROUND:Melanoma differentiation-associated gene-7 (MDA-7)/interleukin-24(IL-24)is a novel tumor suppressor gene,which has suppressor activity in a broad spectrum of human cancer cells.We investigated the effect of the replication-competent oncolytic adenovirus SG600-IL24 and replication-incompetent adenovirus Ad.IL-24,both expressing human MDA-7/IL-24 on the hepatocellular carcinoma cell lines HepG2,Hep3B,SMMC-7721,HCCLM3,and the normal liver cell line L02. METHODS:Hepatocellular carcinoma cell lines and the normal liver cell line were infected with SG600-IL24 and Ad.IL-24.The mRNA and protein expression of MDA-7/IL-24 in infected cells was confirmed by RT-PCR,ELISA,and Western blotting.MTT assay was used to investigate the proliferation effect.Hoechst staining and Annexin-V and PI staining were performed to study the MDA-7/IL-24 gene expressed in HCC cell lines and the normal liver cell line.Flow cytometry was used to analyse the cell cycle. RESULTS:RT-PCR,ELISA and Western blotting confirmed that the exogenous MDA-7/IL-24 gene was highly expressed in cells infected with SG600-IL24.MTT and apoptosis detection indicated that SG600-IL24 induced growth suppression promoted apoptosis,and blocked cancer cell lines in the G2/M phase in hepatocellular carcinoma cell lines but not in the normal liver cell line. CONCLUSIONS:SG600-IL24 selectively induces growth suppression and apoptosis in hepatocellular carcinoma cell lines in vitro but not in the normal liver cell line L02. Compared with Ad.IL-24,SG600-IL24 dramatically enhances antitumor activity in hepatocellular carcinoma cell lines. BACKGROUND: Melanoma differentiation-associated gene-7 (MDA-7) / interleukin-24 (is a novel tumor suppressor gene, which has a suppressor activity in a broad spectrum of human cancer cells. We investigated the effect of the replication -competent oncolytic adenovirus SG600-IL24 and replication-incompetent adenovirus Ad. IL-24, both expressing human MDA-7 / IL-24 on the hepatocellular carcinoma cell lines HepG2, Hep3B, SMMC-7721, HCCLM3, and the normal liver cell line L02. METHODS: Hepatocellular carcinoma cell lines and the normal liver cell line were infected with SG600-IL24 and Ad.IL-24. The mRNA and protein expression of MDA-7 / IL-24 in infected cells was confirmed by RT-PCR, ELISA and Western blotting. MTT assay was used to investigate the proliferation effect. Hoechst staining and Annexin-V and PI staining were performed to study the MDA-7 / IL-24 gene expressed in HCC cell lines and the normal liver cell line. Flow cytometry was used to analyze the cell cycle. RESULTS: RT-PCR, ELISA and Western blotti ng confirmed that the exogenous MDA-7 / IL-24 gene was highly expressed in infected cells with SG600-IL24. MTT and apoptosis detection indicated that SG600-IL24 induced growth suppression promoted apoptosis, and blocked cancer cell lines in the G2 / M phase in hepatocellular carcinoma cell lines but not in the normal liver cell line. CONCLUSIONS: SG600-IL24 selectively induces growth suppression and apoptosis in hepatocellular carcinoma cell lines in vitro but not in the normal liver cell line L02. Compared with Ad.IL-24, SG600-IL24 dramatically enhances antitumor activity in hepatocellular carcinoma cell lines.
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