Immortalization and characterization of normal oral epithelial cells without using HPV and SV40 genes

Abstract

BackgroundAs oral neoplasm often originates from epithelium, an immortalized epithelial cell line could be useful for the research of oral carcinogenesis. Although several oral epithelial cell lines were reported, they were either derived from cancer or immortalized by human papilloma virus or simian virus 40 genes, which have the potential to induce carcinogenesis. Materials and methodsWe established two immortalized cell lines from human oral epithelium by transducing mutant cyclin dependent kinase 4, cyclin D1, and human telomerase reverse transcriptase with or without dominant-negative p53 into primary-cultured normal oral gingival epithelial cells using recombinant lentivirus vectors and named them MOE (mouth-ordinary-epithelium) 1a and MOE1b, respectively. ResultsMOE1 cells could be passaged for nine months or more, and the morphology of the cells did not change in comparison with that of fresh primary-cultured epithelial cells. MOE1 cells did not show epithelial–mesenchymal transition. MOE1b cells retain functional p53 and were considered to have less risk of genomic instabilities. Anchorage-independent growth was not observed in MOE1 cells. The expressions of cancer-associated genes including keratin-17 were not elevated in MOE1 cells, whereas oral cancer-derived HSC-2 cells showed overexpression of them. Furthermore, interleukin (IL)-1β, IL-6, IL-8, tumor necrosis factor-α, matrix metalloproteinase (MMP)-2, and MMP-9 were induced in response to lipopolysaccharide or heat-killed bacterium in MOE1 cells. DiscussionMOE1 cells kept the characteristics of normal epithelial cells without acquiring typical features of cancer cells and they could be useful not only for the study of oral neoplasm but also for other oral diseases.