The induced apoptosis of recombinant adenovirus carrying melittin gene for hepatocellular carcinoma cell / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To observe the induced apoptosis of recombinant adenovirus carrying melittin gene (Ad-rAFP-Mel) for hepatocellular carcinoma cell line (BEL-7402).</p><p><b>METHODS</b>The morphological observe, DNA electrophoresis, TUNEL and Flow cytometry assay were used to study the apoptosis of BEL-7042 cell line transfected by Ad-rAFP-Mel.</p><p><b>RESULTS</b>The morphological changes and apoptosis of BEL-7402 transfected by Ad-rAFP-Mel were confirmed with microscopy and DNA electrophoresis, TUNEL, Flow cytometry assay. The DNA ladder could be demonstrated on DNA electrophoresis in Ad-rAFP-Mel group. The apoptosis rates of BEL-7402 cells in Ad-rAFP-Mel, Ad-rAFP, and control groups were (21.5+/-2.4)%, (10.5+/-4.4)% and (3.0+/-1.4)% respectively by TUNEL assay (F = 38.0, P < 0.05) and were (7.3+/-0.5)%, (3.9+/-0.1)% and (0.8+/-0.1)% respectively by flow cytometry assay (F = 415.1, P < 0.05).</p><p><b>CONCLUSION</b>It seems that melittin inducing apoptosis might be one of the antitumor mechanisms.</p>

Anti-hepatocarcinoma effect of recombinant adenovirus carrying melittin gene / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To find a new method of treating hepatocellular carcinoma with melittin by way of using the melittin gene.</p><p><b>METHODS</b>The recombinant adenoviruses carrying the melittin gene and alpha-fetoprotein (AFP) promoter (Ad-rAFP-Mel) were constructed through a bacterial homologous recombinant system. The efficiency of the adenovirus mediated gene transfer and the inhibition effect of Ad-rAFP-Mel on the proliferation of hepatocarcinoma cells were determined by X-gal staining and MTT assay respectively. The tumorigenicity of hepatocarcinoma cells transfected by Ad-rAFP-Mel and the antitumor effect of Ad-rAFP-Mel on the transplanted tumors in nude mice were detected in vivo.</p><p><b>RESULTS</b>The mRNA of the melittin gene was transcripted in HepG2 hepatocellular carcinoma cells transducted by Ad-rAFP-Mel. The efficiency of adenovirus mediated gene transfered to BEL-7402 hepatocarcinoma cells was 100% when the multiplicities of infection (MOI) of Ad-rAFP-Mel was 10 in vitro and was high in vivo as well. The inhibitive rates of Ad-rAFP-Mel and Ad-rAFP for BEL7402 cells were 66.2%+/-2.7% and 2.9%+/-2.3% (t = 30.83) by MTT assay. The inhibitive rates of Ad-CMV-Mel for BEL7402, SMMC7721 and L02 cells were 58.9%+/-9.6%, 65.9%+/-3.8%, 31.7%+/-1.2%, respectively, and those of the Ad-rAFP-Mel were 6.2%+/-2.7%, 16.1%+/-6.6%, 7.5%+/-3.3%, respectively (t = 1.27; t = 11.31, and t = 12.12, vs. Ad-CMV-Mel group in same cells). The tumorigenicity rates of hepatocarcinoma cells transfected by Ad-rAFP-Mel were decreased. A significant antineoplastic effect was detectd on transplanted tumor in nude mice by intratumoral injection of Ad-rAFP-Mel.</p><p><b>CONCLUSION</b>Ad-rAFP-Mel can inhibit specifically the proliferation of AFP-producing human hepatocarcinoma cells in vitro and in vivo. It suggests that animal toxin gene can be used as an interesting antitumor gene.</p>