1. ESTABLISHMENT AND CHARACTERIZATION OF CELL LINES DERIVED FROM GENETICALLY TRANSFORMED FBM CELLS.
- Author
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Dange, P. P., Kanojia, D., Sawant, S. S., Ingle, A. D., Amare, P. S., and Vaidya, M. M.
- Subjects
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KERATIN , *CELL lines , *EPITHELIAL cells , *CELL transformation , *GENES - Abstract
Keratins (K) are epithelia specific intermediate filament (IF) proteins expressed in a differentiation-dependent and tissue-specific manner. Twenty eight different polypeptides of human keratins have been identified. Keratins are subdivided into two subfamilies, i.e. acidic - with low molecular weights and basic - with high molecular weights, and are always expressed in pairs comprising one member of each subfamily. The keratin pair of 8 and 18 is predominantly expressed by all simple epithelia. This pair is known to be aberrantly expressed by a variety of squamous cell carcinomas. FBM is an immortalized but non-transformed cell line derived in our laboratory from the buccal mucosa of a human fetus. To understand the role of K8/18 in malignant transformation of stratified epithelial cells, full-length keratin 8 gene was transfected in FBM cells. Transfection of K8 gene in these cells resulted in the formation of K8/18 filaments and concomitant malignant transformation of the cells. These cells when injected subcutaneously in nude mice, produced tumors at the site of injection. Three tumor cell lines have been established from three tumors derived from three different K8 transected FBM clones. These cell lines are being further characterized. Keratin expression pattern, cell doubling time, soft agar colony forming assay, cell cycle analysis using flow cytometry, growth supplement independence and nude mice injections were carried out. Analysis of keratin expression based on 2-D gel electrophoresis followed by western blot has demonstrated expression of keratins 1, 5, 7, 10 in one of the cell lines. Keratin 8/18 expression was seen in all the cell lines similar to transfected clones. These cell lines showed reduced doubling time compared to parental K8 transfected clones, anchorage independent growth and tumors in nude mice. These cells also showed polyploidy. These cell lines will be useful to study different aspects of oral carcinogenesis, including chemoprevention. [ABSTRACT FROM AUTHOR]
- Published
- 2007