Experimental Study on Analgesic-antitumor Peptide from Buthus martensii Karsch Targeting to Hepatocellular Carcinoma in vitro

Background:Hepatocellular carcinoma(HCC) is one of the most common malignancies worldwide;however,the efficacy of conventional therapies such as surgical operation,interventional therapy and radiochemotherapy is not so satisfactory.New treatment modalities with higher efficacy and lower toxic reaction should be pursued.Aims:To assess the targeting and cytotoxic effects of eukaryotic expression vector carrying analgesic-antitumor peptide(AGAP) gene derived from Buthus martensii Karsch on HCC cells.Methods:Eukaryotic expression plasmid pAFP-AGAP was identified by enzyme digestion and sequencing,and then transfected intoα-fetoprotein(AFP)-positive human HCC cell line HepG2 and AFP-negative human cervical carcinoma cell line HeLa and normal human hepatic cell line L02.The expression level of AGAP mRNA was determined by reverse transcriptase polymerase chain reaction(RT-PCR),and Cell Counting Kit-8 was used to analyze the cytotoxicity.Results:AGAP mRNA was detected in AFP-producing HepG2 cells but not in HeLa and L02 cells after plasmid pAFP-AGAP transfection.Forty-eight hours after transfection with plasmid pAFP-AGAP, significant morphological changes and growth inhibition(P0.01) were observed in HepG2 cells,the inhibition rate at 48- hour and 72-hour was 44.4%and 74.6%,respectively;whereas no such changes were found in HeLa and L02 cells. Conclusions:AGAP gene eukaryotic expression vector can selectively target the HCC cells with high cytotoxic efficacy, which might be a promising effective tool for HCC gene therapy.