Two-dimensional electrophoretic studies on down-regulated intracellular transferrin in human fibroblasts immortalized by treatment with either 4-nitroquinoline 1-oxide or 60Co gamma rays.

The immortalization of human cells is an important early step of carcinogenesis. To investigate the mechanisms of immortalization, we established three immortalized human fibroblast cell lines by treatment with either 4-nitroquinoline 1-oxide (4NQO) or 60Co gama rays. Using two-dimensional gel electrophoresis (2-D PAGE), we identified proteins which are down-regulated in these immortalization cell lines. Their isoelectric points ranged between 5.5 and 6.3, and their molecular masses were approximately 80 kDa. The proteins were also decreased in another human fibroblast cell line immortalized with simian virus 40 (SV40) and two human cervical cell lines. The proteins were present in a cytoskeletal fraction, reacted with anti-transferrin antibody and some of them were phosphorylated. The proteins were identified as transferrin. Although the 2-D PAGE patterns of our proteins and serum transferrin of different origin were distinct from each other, their isoelectric points and molecular weights were similar. We implicate some functions of intracellular transferrin, other than an iron transporter, in immortalization of human cells and discuss a new aspect of transferrin metabolism in the cells.