Effects of liposome-mediated rat cyclooxygenase-2 antisense RNA on growth of hepatocarcinoma cells

Abstract

Objective To construct a recombinant eukaryotic expression vector encoding rat COX-2 antisense RNA and investigate its effects on rat liver cancer cell proliferatiion. Methods The plasmid encoding anti-sense COX-2 was constructed by using cloning COX-2 cDNA fragment in the reverse direction into the pcDNA3. 1. Then the plasmid pcDNA3. l/COX-2as was transfered into rat hepatocacinoma cell line CBRH7919 with liposome and homologous recombination cell was named as CBRH7919-A. The cell proliferation, cell cycle and apoptosis were analyzed by MTT, flow cytometry and RT-PCR. CBRH7919 cells with or without pretreatment were inoculated into nude mice and the cell proliferation was observed in vivo. Results CBRH7919 treated with antisense COX-2 greatly inhibited the proliferation with a rate of 78% and DNA synthesis (PI of COX-2 group vs control group, 0. 361 vs 0. 784) of CBRH7919 cell line, decreased coloning formation in anchorage-independent assay. In vivo tumorigenic rate was greatly reduced in athymic nude mice as compared with untreated cell group (25% vs 100%). Apoptosis-related gene expression was not different as compared with untreated cell group (P>0. 05). Conclusion Antisense COX-2 RNA can inhibit the proliferation of rat liver cancer cell line in vitro as well as in vivo, suggesting that it has potential value in hepatocellular cancer gene therapy. Key words: Carcinoma hepatocellular; Liposome; Rat; Cyclooxygenase-2; Antisense RNA