Establishment and characterization of a cell line from smokeless tobacco associated oral squamous cell carcinoma.

A cell line, AMOS-III has been established from the surgically resected specimen of an untreated primary human oral squamous cell carcinoma of the floor of mouth from a chronic smokeless tobacco consumer. Immunocytochemical analysis showed epithelial specific antigen, cytokeratins 5, 10, 13 and 16 and integrin alpha(6) markers in AMOS-III cells, confirming the epithelial lineage of the cell line. Analyses of morphology, ultrastructure, karyotype, anchorage independent growth and immunocytochemical properties of the cell line demonstrated the transformed phenotype of epithelial cells. AMOS-III cells have doubling time of 42-44 h. Giemsa-banding patterns of chromosomes confirmed the human origin of the AMOS-III cells. Molecular analysis of cancer-related gene products, p53 and p21(cip1/waf1) showed the presence of wild type p21(cip1/waf1) and truncated p53 proteins. The molecular mechanism underlying the action of retinoids in preventing the occurrence of second primary tumors in oral cancer patients remain to be clearly defined. Treatment of AMOS-III cells with all-trans retinoic acid (ATRA) at 10(-4) microM resulted in 81% cell death. ATRA treatment resulted in enhanced expression of p21(cip1/waf1), nuclear translocation of retinoic acid receptors and apoptotic cell death. Thus, this cell line provides an in vitro model for elucidating the mechanism involving p53 inactivation and p21(cip1/waf1) overexpression in smokeless tobacco-induced oral cancer. Furthermore, the ATRA responsiveness of the cell line underscores its potential utility in identifying the retinoid responsive molecular targets in oral cancer cells.