The expression of therapeutic gene and its anti-tumor effects will beaugmented and a synergism of oncolytic virus with the therapeutic gene is speculated. This study wasundertaken to assess the anti-tumor effects of a...The expression of therapeutic gene and its anti-tumor effects will beaugmented and a synergism of oncolytic virus with the therapeutic gene is speculated. This study wasundertaken to assess the anti-tumor effects of a novel gene-viral therapeutic systemCNHK300-mEndostatin ( CNHK300-mE) in hepatocellular carcinoma (HCC). A novel gene-viral therapeuticsystem named CNHK300-mE was constructed using the human telomerase reverse transcriptase (hTERT)promoter to drive the expression of the adenovirus El A gene and cloning the therapeutic gene mouseendostatin into the adenovirus genome. By the tissue culture infectious dose 50 (TCID50) method andcytoviability assay, the replicative and cytolytic capabilities of CNHK300-mE in two HCC lines (HepGII and Hep3B) and one normal cell line ( MRC-5 ) were analyzed, and the transgene expressions ofmouse endostatin in vitro and in vivo were detected by Western blotting and ELISA assay. Tumorgrowth suppression and anti-angiogenesis effects in vivo were investigated using nude micexenografts model derived from SMMC-7721 HCC cells. The 3296-fold replicating capacity of CNHK300-mEin HCC cell lines versus in the normal cell line at 96 hours post infection and the 25 -foldeffective dose for killing 50% cells ( ED50) in the normal cell line versus HCC cell lines, whichwere both superior to ONYX-015, were observed. Tumor growth suppression of CNHK300-mE superior toeither Ad-mE or ONYX-015 was demonstrated (P < 0. 01) and the anti-angiogenic effects in vivosuperior to Ad-mE were also observed with immunohistochemical staining of von Willebrand factor. Incomparison with non-replicative adenovirus Ad-mE, the transgene expression of mE mediated byCNHK300- in vivo (P<0.05). Being capable of replicating in and lysing the telomerase-positive HCCcells and mediating effective expression of the therapeutic gene in vitro and in vivo, the novelgene-viral therapeutic system CNHK300-mE is potentially effective in the treatment of HCC.展开更多
文摘The expression of therapeutic gene and its anti-tumor effects will beaugmented and a synergism of oncolytic virus with the therapeutic gene is speculated. This study wasundertaken to assess the anti-tumor effects of a novel gene-viral therapeutic systemCNHK300-mEndostatin ( CNHK300-mE) in hepatocellular carcinoma (HCC). A novel gene-viral therapeuticsystem named CNHK300-mE was constructed using the human telomerase reverse transcriptase (hTERT)promoter to drive the expression of the adenovirus El A gene and cloning the therapeutic gene mouseendostatin into the adenovirus genome. By the tissue culture infectious dose 50 (TCID50) method andcytoviability assay, the replicative and cytolytic capabilities of CNHK300-mE in two HCC lines (HepGII and Hep3B) and one normal cell line ( MRC-5 ) were analyzed, and the transgene expressions ofmouse endostatin in vitro and in vivo were detected by Western blotting and ELISA assay. Tumorgrowth suppression and anti-angiogenesis effects in vivo were investigated using nude micexenografts model derived from SMMC-7721 HCC cells. The 3296-fold replicating capacity of CNHK300-mEin HCC cell lines versus in the normal cell line at 96 hours post infection and the 25 -foldeffective dose for killing 50% cells ( ED50) in the normal cell line versus HCC cell lines, whichwere both superior to ONYX-015, were observed. Tumor growth suppression of CNHK300-mE superior toeither Ad-mE or ONYX-015 was demonstrated (P < 0. 01) and the anti-angiogenic effects in vivosuperior to Ad-mE were also observed with immunohistochemical staining of von Willebrand factor. Incomparison with non-replicative adenovirus Ad-mE, the transgene expression of mE mediated byCNHK300- in vivo (P<0.05). Being capable of replicating in and lysing the telomerase-positive HCCcells and mediating effective expression of the therapeutic gene in vitro and in vivo, the novelgene-viral therapeutic system CNHK300-mE is potentially effective in the treatment of HCC.
