TRANSFORMATION OF HUMAN KERATINOCYTES IS CHARACTERIZED BY QUANTITATIVE AND QUALITATIVE ALTERATIONS OF THE T-16 ANTIGEN (TROP-2, MOV-16)

The regulation of synthesis and post-translational processing of the T-16 antigen, a human cell-surface glycoprotein of 50 to 60 kDa, was investigated in normal and transformed human keratinocytes in vitro. Normal keratinocytes of follicular origin were compared with squamous-cell-carcinoma lines, spontaneously immortalized keratinocytes, and SV-40 transformed keratinocytes. FAGS analysis and radioimmunoprecipitation showed that the synthesis and expression of T-16 was 3- to 4-fold higher in transformed keratinocytes than in their normal counterparts. In normal keratinocytes, no quantitative differences were observed among freshly prepared cells, primary cultures and sub-cultures. In SDS-PAGE, a single broad band at 50 to 60 kDa was observed in normal keratinocytes, whereas 2 bands at 42 and 45 to 55 kDa were detected after transformation. Tunicamycin treatment of living cells and glycosidase digestion of immunopurified T-16 antigen revealed this molecular heterogeneity to be due to different N-glycosylation in normal and transformed keratinocytes. In pulse-chase experiments, 2 distinct precursor proteins at 38 and 42 kDa were detected in transformed keratinocytes, whereas in normal cells the 38-kDa signal was dramatically decreased. These findings indicate that quantitative and qualitative changes of T-16 mark the transformation process of human keratinocytes, showing similar post-translational alterations in all transformed populations investigated. (C) 1995 Wiley-Liss, Inc.