Expression of neuroendocrine and epithelial markers in an adherent subline derived from a classic small cell lung cancer cell line

Abstract

Small cell lung cancer (SCLC) cell lines usually grow as floating aggregates, in contrast to the adherent monolayers formed by non small cell lung cancer (NSCLC). Induction of an adherent phenotype by a variety of methods has been the subject of a number of recent publications. In this study, cultivation of the classic SCLC cell line, NCI-H69, on a substratum provided by the pretreatment of tissue culture dishes with medium conditioned by the growth of a well differentiated squamous carcinoma cell line, HN5, induced an adherent phenotype with a variety of epithelioid morphologies, commencing within 24 hr of plating. From such cultures an adherent subline, H69A, has been established, which differs in its growth, morphological characteristics, and immunocytochemical marker expression from the parent NCI-H69 cells, and in its marker expression from other adherent SCLC cell lines. H69A retained expression of neural cell adhesion molecule (NCAM) and the neuroendocrine markers neuron specific enoclase, chromogranin A, and synaptophysin, but showed diminished expression of the epithelial cell surface markers AUA1, Ber-EP4, epithelial membrane antigen (EMA), and desmosomal protein. Compared to NCI-H69 cells, the amounts of cytokeratin 18 detected were elevated, while those of cytokeratin 19 were diminished in H69A cells. Focal expression of cytokeratin 4 was found in some H69A cells, indicative of a capacity for partial squamous differentiation. The expression of the cell surface glycoproteins detected by AUA1 and Ber-EP4 was reduced throughout cultivation of the H69A subline, while that of EMA and desmosomal protein was further diminished with continued passage. Changes in the expression of these markers and NCAM were evident in NCI-H69 cells grown on an HN5-derived substratum.