Expression of PSA-N-CAM in human neuroblastoma cells induced to neuronal differentiation by retinoic acid.

The neural cell adhesion molecule (N-CAM) plays a significant role in the development of the nervous system. Three different isoforms of the molecule have been described, with molecular masses of 180, 140 and 120 kDa, whose differential expression in neurons seems to be related to their state of differentiation. We took advantage of the use of the human neuroblastoma cell line LAN-5, which can be differentiated in vitro by retinoic acid (RA) into neuronal cells, for studying the expression of N-CAM isoforms, and their polysialic acid (PSA) content, at the protein and mRNA levels. Anti-N-CAM polyclonal antibodies recognizing all the N-CAM isoforms and a monoclonal antibody recognizing PSA were used in Western blot experiments with extracts from undifferentiated and RA-differentiated cells. We found that undifferentiated cells express very little of the 180 kDa N-CAM isoform and a large amount of the 140 kDa isoform. A 4-fold increase in the expression of the 180 kDa N-CAM isoform was obtained when LAN-5 cells were differentiated by RA for 8 days, whereas a 1.8-fold increase in the expression of the 140 kDa N-CAM isoform was observed upon differentiation. Similarly, the levels of the 7.4 kb mRNA coding for N-CAM 180 kDa, determined by Northern blot analysis, were barely detectable in undifferentiated cells, and showed a 3.8-fold increase upon differentiation. By contrast, only a 1.3-fold increase in the 6.7 kb mRNA, coding for the 140 kDa N-CAM isoform, was observed. N-CAM was always found in its polysialylated form in both undifferentiated and RA-differentiated cells. This indicates that, in LAN-5 cells, the expression and activity of the polysialytransferase enzyme precedes the acquisition of a neuronal phenotype.