Stable long term transfected human hepatocellular carcinoma cell line bearing HSV 1 thymidine kinase gene by retroviral vector

The recombinant retroviral vector pLTRNL containing the HSV 1 TK gene was transfected into human hepatocellular carcinoma cell line hHCC by lipofectamine method. The transfected cells were selected with 800mg/L of geneticin(G418) for three weeks, and then cloned and subcloned. G418 resistant transfected cells were named hHCC/TK cells. The total RNA and genomic DNA of hHCC/TK or hHCC were extracted. These genomic DNA containing TK gene were identified by PCR. The total RNA containing mRNA of HSV 1 TK gene were identified by RT PCR and RNA slot blot methods. The results showed that the HSV 1 TK gene was integrated into the genomc of transfected cells and the mRNA of TK gene has been transcribed. The establishment of stable long term transfected human hepatocellular carcinoma cell line bearing HSV 1 TK gene could provide the possibility of experimental gene therapy for human hepatocellular carcinoma in vitro and in vivo .