ObjectiveTo construct a hepatocellular carcinoma (HCC) cell line stably transfected with linamarase (lis) gene from cassava, and to study its biological characteristics. MethodsLis DNA was amplified from cassava by PCR and cloned into the pcDNA3.1(+) plasmid. Then the recombinant plasmid pcDNA3.1/lis was transfected into human HCC HepG2 cell line using lipofectamineTM 2000 and G418 selection. The stably transfected cell lines HepG2/lis was identified by immunofluorescence, RT-PCR, and Western blot. The enzyme activity of lis in cells was assayed by Lambert method. The characteristics of the new cell lines were checked by several methods: the cell growth curve was observed by MTT, cell cycle was analyzed by flow cytometry, and characteristics of tumor formation in vivo were detected in nude mice. ResultsLis from the cassava could stably integrate into eukaryotic cells, and package the same protein as lis with β-glucosidase activity. The stable integration of lis in cells did not interfere with cell morphology, growth characteristics, cell cycle, and tumorigenesis in vivo significantly. ConclusionsA new HCC cell lines transfected with lis is successfully established, which may lay an experimental foundation for the study of HCC treatment by using lis suicide gene system in future.