Construction of eukaryotic vector expressing antisense RNA of hepatoma associated antigen-HAb18G and clone identification

AIM To construct eukaryotic vector expressing antisense RNA of HAb18G. METHODS PCI asHAb18G was constructed by inserting HAb18G cDNA reversely to eukaryotic expression vector PCI neo. The hepatoma cell strain HHCC was transfected by PCI asHAb18G. After selected byG418, positive clones were identified by fluorocytometry assay and indirect immunofluorescence staining. RESULTS It was verified that construction of the eukaryotic antisense RNA expression vector PCI asHAb18G was correct by partial nucleotide sequencing and restriction endonuclease digestion. Indirect immunofluorescence staining and fluorocytometric assay show that the expression of HAb18G was negtive in HHCC transfected by PCI asHAb18G and its expression was positive in HHCC transfected by PCI neo and untransfected HHCC. CONCLUSION These results lay the foundation for further study of the biological functions of HAb18G /CD147 and gene therapy for hepatoma.