Objective An adeno associated virus (AAV)vector targeted to hepatoma cells was constructed in order to be used in the apoptosis inducing gene therapy of liver cancer. Methods Firstly, primers containing specific enzyme cutting sites was designed and used to amplify the alpha fetoprotein promoter(AFP promoter) from human genome; Secondly, the promoter was cloned into the eukyrocyte expressing plasmid pTR UF5,resulting in the recombinant AAV vector plasmid containing the reporter gene( rAAV AFP GFP ); thirdly, blunted ligation was applied to construct the recombinant AAV vector plasmid containing the p53 gene( rAAV AFP p53 ). Results Recombinant adeno associated virus plasmid was constructed successfully that carried human wild type p53 gene under the control of human AFP promoter. Conclusion Theoretically the recombinant adeno associated virus plasmid rAAV AFP p53 we constructed could be used in gene therapy for hepatocellular carcinoma.

Abstract:Objective To detect the mutation frequency of the bcl 10 gene in the early and advanced stages of hepatocellular carcinoma (HCC).Methods Genome DNA samples were extracted from 46 cases of fresh HCC tumor tissues and their non-tumor adjacent tissues. Polymerase chain reaction-single strand conformation polymorphism method was used to detect point mutations of the three exons of the bcl 10 gene. For each individual exon, six random samples from those showing abnormal DNA bands were sequenced to verify those mutations. The relationship between serum alpha-fetoprotein (AFP) level and bcl 10 mutation, between the tumor size and bcl 10 mutation was also analyzed.Results Among the 46 samples, 26 cases (56.5%) were found to have mutations in exon 1, 5 out of the 6 cases were shown to have 5744 C→G mutation by sequencing; 25 cases (54.3%) were found to have mutations in exon 2, 4 out of the 6 cases were shown to have 11?311 T deletion mutation by sequencing. Twenty-one cases (45.7%) were found to have mutations in exon 3, all of the 6 cases selected for sequencing were shown to have 14?116 C→T mutation. Statistical analysis showed that neither serum alpha-fetoprotein level nor the size of hepatocellular carcinoma has a significant relationship with bcl 10 mutation.Conclusion The bcl 10 gene has a high mutation frequency in liver cancer.